Difference between revisions of "Protocol.html"

Line 10: Line 10:
 
<meta name=Generator content="Microsoft Word 14">
 
<meta name=Generator content="Microsoft Word 14">
 
<meta name=Originator content="Microsoft Word 14">
 
<meta name=Originator content="Microsoft Word 14">
<link rel=File-List href="PROToCOLS_files/filelist.xml">
+
<link rel=File-List href="INTER_files/filelist.xml">
<link rel=Edit-Time-Data href="PROToCOLS_files/editdata.mso">
+
<link rel=Edit-Time-Data href="INTER_files/editdata.mso">
 
<!--[if !mso]>
 
<!--[if !mso]>
 
<style>
 
<style>
Line 24: Line 24:
 
   <o:LastAuthor>hp</o:LastAuthor>
 
   <o:LastAuthor>hp</o:LastAuthor>
 
   <o:Revision>1</o:Revision>
 
   <o:Revision>1</o:Revision>
   <o:TotalTime>8</o:TotalTime>
+
   <o:TotalTime>2</o:TotalTime>
   <o:Created>2017-11-01T20:38:00Z</o:Created>
+
   <o:Created>2017-11-01T21:25:00Z</o:Created>
   <o:LastSaved>2017-11-01T20:50:00Z</o:LastSaved>
+
   <o:LastSaved>2017-11-01T21:27:00Z</o:LastSaved>
   <o:Pages>2</o:Pages>
+
   <o:Pages>1</o:Pages>
   <o:Words>340</o:Words>
+
   <o:Words>2</o:Words>
   <o:Characters>1941</o:Characters>
+
   <o:Characters>16</o:Characters>
   <o:Lines>16</o:Lines>
+
   <o:Lines>1</o:Lines>
   <o:Paragraphs>4</o:Paragraphs>
+
   <o:Paragraphs>1</o:Paragraphs>
   <o:CharactersWithSpaces>2277</o:CharactersWithSpaces>
+
   <o:CharactersWithSpaces>17</o:CharactersWithSpaces>
 
   <o:Version>14.00</o:Version>
 
   <o:Version>14.00</o:Version>
 
  </o:DocumentProperties>
 
  </o:DocumentProperties>
Line 39: Line 39:
 
  </o:OfficeDocumentSettings>
 
  </o:OfficeDocumentSettings>
 
</xml><![endif]-->
 
</xml><![endif]-->
<link rel=dataStoreItem href="PROToCOLS_files/item0001.xml"
+
<link rel=dataStoreItem href="INTER_files/item0001.xml"
target="PROToCOLS_files/props002.xml">
+
target="INTER_files/props002.xml">
<link rel=themeData href="PROToCOLS_files/themedata.thmx">
+
<link rel=themeData href="INTER_files/themedata.thmx">
<link rel=colorSchemeMapping href="PROToCOLS_files/colorschememapping.xml">
+
<link rel=colorSchemeMapping href="INTER_files/colorschememapping.xml">
 
<!--[if gte mso 9]><xml>
 
<!--[if gte mso 9]><xml>
 
  <w:WordDocument>
 
  <w:WordDocument>
Line 345: Line 345:
 
  /* Font Definitions */
 
  /* Font Definitions */
 
  @font-face
 
  @font-face
{font-family:Wingdings;
 
panose-1:5 0 0 0 0 0 0 0 0 0;
 
mso-font-charset:2;
 
mso-generic-font-family:auto;
 
mso-font-pitch:variable;
 
mso-font-signature:0 268435456 0 0 -2147483648 0;}
 
@font-face
 
{font-family:Wingdings;
 
panose-1:5 0 0 0 0 0 0 0 0 0;
 
mso-font-charset:2;
 
mso-generic-font-family:auto;
 
mso-font-pitch:variable;
 
mso-font-signature:0 268435456 0 0 -2147483648 0;}
 
@font-face
 
 
{font-family:Calibri;
 
{font-family:Calibri;
 
panose-1:2 15 5 2 2 2 4 3 2 4;
 
panose-1:2 15 5 2 2 2 4 3 2 4;
Line 365: Line 351:
 
mso-font-pitch:variable;
 
mso-font-pitch:variable;
 
mso-font-signature:-520092929 1073786111 9 0 415 0;}
 
mso-font-signature:-520092929 1073786111 9 0 415 0;}
@font-face
 
{font-family:Tahoma;
 
panose-1:2 11 6 4 3 5 4 4 2 4;
 
mso-font-charset:0;
 
mso-generic-font-family:swiss;
 
mso-font-pitch:variable;
 
mso-font-signature:-520081665 -1073717157 41 0 66047 0;}
 
@font-face
 
{font-family:"Book Antiqua";
 
panose-1:2 4 6 2 5 3 5 3 3 4;
 
mso-font-charset:0;
 
mso-generic-font-family:roman;
 
mso-font-pitch:variable;
 
mso-font-signature:647 0 0 0 159 0;}
 
@font-face
 
{font-family:Algerian;
 
panose-1:4 2 7 5 4 10 2 6 7 2;
 
mso-font-charset:0;
 
mso-generic-font-family:decorative;
 
mso-font-pitch:variable;
 
mso-font-signature:3 0 0 0 1 0;}
 
@font-face
 
{font-family:"Open Sans";
 
panose-1:0 0 0 0 0 0 0 0 0 0;
 
mso-font-alt:"Times New Roman";
 
mso-font-charset:0;
 
mso-generic-font-family:roman;
 
mso-font-format:other;
 
mso-font-pitch:auto;
 
mso-font-signature:0 0 0 0 0 0;}
 
 
  /* Style Definitions */
 
  /* Style Definitions */
 
  p.MsoNormal, li.MsoNormal, div.MsoNormal
 
  p.MsoNormal, li.MsoNormal, div.MsoNormal
Line 416: Line 372:
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;}
 
mso-bidi-theme-font:minor-bidi;}
a:link, span.MsoHyperlink
 
{mso-style-priority:99;
 
color:blue;
 
text-decoration:underline;
 
text-underline:single;}
 
a:visited, span.MsoHyperlinkFollowed
 
{mso-style-noshow:yes;
 
mso-style-priority:99;
 
color:purple;
 
mso-themecolor:followedhyperlink;
 
text-decoration:underline;
 
text-underline:single;}
 
p
 
{mso-style-priority:99;
 
mso-margin-top-alt:auto;
 
margin-right:0in;
 
mso-margin-bottom-alt:auto;
 
margin-left:0in;
 
mso-pagination:widow-orphan;
 
font-size:12.0pt;
 
font-family:"Times New Roman","serif";
 
mso-fareast-font-family:"Times New Roman";
 
mso-fareast-theme-font:minor-fareast;}
 
p.MsoAcetate, li.MsoAcetate, div.MsoAcetate
 
{mso-style-noshow:yes;
 
mso-style-priority:99;
 
mso-style-link:"Balloon Text Char";
 
margin:0in;
 
margin-bottom:.0001pt;
 
mso-pagination:widow-orphan;
 
font-size:8.0pt;
 
font-family:"Tahoma","sans-serif";
 
mso-fareast-font-family:Calibri;
 
mso-fareast-theme-font:minor-latin;}
 
p.MsoListParagraph, li.MsoListParagraph, div.MsoListParagraph
 
{mso-style-priority:34;
 
mso-style-unhide:no;
 
mso-style-qformat:yes;
 
margin-top:0in;
 
margin-right:0in;
 
margin-bottom:10.0pt;
 
margin-left:.5in;
 
mso-add-space:auto;
 
line-height:115%;
 
mso-pagination:widow-orphan;
 
font-size:11.0pt;
 
font-family:"Calibri","sans-serif";
 
mso-ascii-font-family:Calibri;
 
mso-ascii-theme-font:minor-latin;
 
mso-fareast-font-family:Calibri;
 
mso-fareast-theme-font:minor-latin;
 
mso-hansi-font-family:Calibri;
 
mso-hansi-theme-font:minor-latin;
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;}
 
p.MsoListParagraphCxSpFirst, li.MsoListParagraphCxSpFirst, div.MsoListParagraphCxSpFirst
 
{mso-style-priority:34;
 
mso-style-unhide:no;
 
mso-style-qformat:yes;
 
mso-style-type:export-only;
 
margin-top:0in;
 
margin-right:0in;
 
margin-bottom:0in;
 
margin-left:.5in;
 
margin-bottom:.0001pt;
 
mso-add-space:auto;
 
line-height:115%;
 
mso-pagination:widow-orphan;
 
font-size:11.0pt;
 
font-family:"Calibri","sans-serif";
 
mso-ascii-font-family:Calibri;
 
mso-ascii-theme-font:minor-latin;
 
mso-fareast-font-family:Calibri;
 
mso-fareast-theme-font:minor-latin;
 
mso-hansi-font-family:Calibri;
 
mso-hansi-theme-font:minor-latin;
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;}
 
p.MsoListParagraphCxSpMiddle, li.MsoListParagraphCxSpMiddle, div.MsoListParagraphCxSpMiddle
 
{mso-style-priority:34;
 
mso-style-unhide:no;
 
mso-style-qformat:yes;
 
mso-style-type:export-only;
 
margin-top:0in;
 
margin-right:0in;
 
margin-bottom:0in;
 
margin-left:.5in;
 
margin-bottom:.0001pt;
 
mso-add-space:auto;
 
line-height:115%;
 
mso-pagination:widow-orphan;
 
font-size:11.0pt;
 
font-family:"Calibri","sans-serif";
 
mso-ascii-font-family:Calibri;
 
mso-ascii-theme-font:minor-latin;
 
mso-fareast-font-family:Calibri;
 
mso-fareast-theme-font:minor-latin;
 
mso-hansi-font-family:Calibri;
 
mso-hansi-theme-font:minor-latin;
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;}
 
p.MsoListParagraphCxSpLast, li.MsoListParagraphCxSpLast, div.MsoListParagraphCxSpLast
 
{mso-style-priority:34;
 
mso-style-unhide:no;
 
mso-style-qformat:yes;
 
mso-style-type:export-only;
 
margin-top:0in;
 
margin-right:0in;
 
margin-bottom:10.0pt;
 
margin-left:.5in;
 
mso-add-space:auto;
 
line-height:115%;
 
mso-pagination:widow-orphan;
 
font-size:11.0pt;
 
font-family:"Calibri","sans-serif";
 
mso-ascii-font-family:Calibri;
 
mso-ascii-theme-font:minor-latin;
 
mso-fareast-font-family:Calibri;
 
mso-fareast-theme-font:minor-latin;
 
mso-hansi-font-family:Calibri;
 
mso-hansi-theme-font:minor-latin;
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;}
 
span.BalloonTextChar
 
{mso-style-name:"Balloon Text Char";
 
mso-style-noshow:yes;
 
mso-style-priority:99;
 
mso-style-unhide:no;
 
mso-style-locked:yes;
 
mso-style-link:"Balloon Text";
 
mso-ansi-font-size:8.0pt;
 
mso-bidi-font-size:8.0pt;
 
font-family:"Tahoma","sans-serif";
 
mso-ascii-font-family:Tahoma;
 
mso-hansi-font-family:Tahoma;
 
mso-bidi-font-family:Tahoma;}
 
span.SpellE
 
{mso-style-name:"";
 
mso-spl-e:yes;}
 
span.GramE
 
{mso-style-name:"";
 
mso-gram-e:yes;}
 
 
.MsoChpDefault
 
.MsoChpDefault
 
{mso-style-type:export-only;
 
{mso-style-type:export-only;
Line 582: Line 396:
 
div.WordSection1
 
div.WordSection1
 
{page:WordSection1;}
 
{page:WordSection1;}
/* List Definitions */
 
@list l0
 
{mso-list-id:17397210;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-154127740 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
 
@list l0:level1
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l0:level2
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l0:level3
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l0:level4
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l0:level5
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l0:level6
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l0:level7
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l0:level8
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l0:level9
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l1
 
{mso-list-id:527305042;
 
mso-list-type:hybrid;
 
mso-list-template-ids:1506328854 -268911842 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l1:level1
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Times New Roman","serif";
 
mso-ascii-font-family:Calibri;
 
mso-ascii-theme-font:minor-latin;
 
mso-hansi-font-family:Calibri;
 
mso-bidi-font-family:"Times New Roman";
 
mso-bidi-theme-font:minor-bidi;
 
color:black;
 
mso-themecolor:text1;
 
mso-ansi-font-weight:bold;}
 
@list l1:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l1:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l1:level4
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l1:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l1:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l1:level7
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l1:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l1:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l2
 
{mso-list-id:869534640;
 
mso-list-type:hybrid;
 
mso-list-template-ids:366508600 67698703 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l2:level1
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l2:level4
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l2:level7
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l2:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-9.0pt;}
 
@list l3
 
{mso-list-id:938374804;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-786953526 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
 
@list l3:level1
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l3:level2
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l3:level3
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l3:level4
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l3:level5
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l3:level6
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l3:level7
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l3:level8
 
{mso-level-number-format:bullet;
 
mso-level-text:o;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:"Courier New";}
 
@list l3:level9
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
ol
 
{margin-bottom:0in;}
 
ul
 
{margin-bottom:0in;}
 
 
-->
 
-->
 
</style>
 
</style>
Line 851: Line 425:
 
</style>
 
</style>
 
<![endif]--><!--[if gte mso 9]><xml>
 
<![endif]--><!--[if gte mso 9]><xml>
  <o:shapedefaults v:ext="edit" spidmax="1026"/>
+
  <o:shapedefaults v:ext="edit" spidmax="1027"/>
 
</xml><![endif]--><!--[if gte mso 9]><xml>
 
</xml><![endif]--><!--[if gte mso 9]><xml>
 
  <o:shapelayout v:ext="edit">
 
  <o:shapelayout v:ext="edit">
Line 858: Line 432:
 
</head>
 
</head>
  
<body lang=EN-US link=blue vlink=purple style='tab-interval:.5in'>
+
<body lang=EN-US style='tab-interval:.5in'>
  
 
<div class=WordSection1>
 
<div class=WordSection1>
  
 
<p class=MsoNormal align=center style='text-align:center'><span
 
<p class=MsoNormal align=center style='text-align:center'><span
style='font-size:72.0pt;line-height:115%;font-family:Algerian'>PROToCOLS<o:p></o:p></span></p>
+
style='font-size:56.0pt;line-height:115%'>INTER-LAB STUDY<o:p></o:p></span></p>
 
+
<p align=center style='margin-top:0in;margin-right:0in;margin-bottom:0in;
+
margin-left:.5in;margin-bottom:.0001pt;text-align:center;text-indent:-.25in;
+
mso-list:l1 level1 lfo1'><![if !supportLists]><b style='mso-bidi-font-weight:
+
normal'><span style='font-size:14.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;color:black;
+
mso-themecolor:text1;mso-font-kerning:12.0pt'><span style='mso-list:Ignore'>1.<span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span></b><![endif]><b><span
+
style='font-size:20.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'>Protocol for Cloning
+
Experiments</span></b><span style='font-size:14.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
+
mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:text1;mso-font-kerning:
+
12.0pt'><o:p></o:p></span></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify;text-indent:-.25in;line-height:150%;
+
mso-list:l0 level1 lfo4'><![if !supportLists]><span style='font-size:14.0pt;
+
line-height:150%;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
+
Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span style='font-size:14.0pt;line-height:150%;
+
font-family:"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>Protocol for PCR Amplification, Competent Cell
+
Preparation, and Transformation was followed as given in Molecular Cloning: A
+
Laboratory Manual, 3<span class=SpellE><sup><span style='position:relative;
+
top:-8.5pt;mso-text-raise:8.5pt'>rd</span></sup></span><span
+
style='mso-spacerun:yes'> </span>edition Sam brook.</span><span
+
style='font-size:14.0pt;line-height:150%'><o:p></o:p></span></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify;text-indent:-.25in;line-height:150%;
+
mso-list:l0 level1 lfo4'><![if !supportLists]><span style='font-size:14.0pt;
+
line-height:150%;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
+
Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span style='font-size:14.0pt;line-height:150%;
+
font-family:"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>All Restriction Endonuclease used are of Thermo
+
Fisher Scientific and T4DNA ligation Enzyme is of New England Bio labs and all
+
reactions are set as per guidelines of the supplier.</span><span
+
style='font-size:14.0pt;line-height:150%'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt'><span style='font-size:14.0pt'><o:p>&nbsp;</o:p></span></p>
+
 
+
<p align=center style='margin-top:0in;margin-right:0in;margin-bottom:0in;
+
margin-left:.5in;margin-bottom:.0001pt;text-align:center;text-indent:-.25in;
+
mso-list:l1 level1 lfo1'><![if !supportLists]><b style='mso-bidi-font-weight:
+
normal'><span style='font-size:20.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;color:black;
+
mso-themecolor:text1'><span style='mso-list:Ignore'>2.<span style='font:7.0pt "Times New Roman"'>&nbsp;
+
</span></span></span></b><![endif]><b><span style='font-size:20.0pt;font-family:
+
"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>Promoter Characterization</span></b><span
+
style='font-size:20.0pt'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt;text-align:justify;line-height:150%'><span
+
style='font-size:14.0pt;line-height:150%;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
+
mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:text1;mso-font-kerning:
+
12.0pt'>We had done in <i>In-Silico</i> Characterization of Promoter by
+
validating the <i style='mso-bidi-font-style:normal'>cis-</i>element of the
+
promoters by using Plant Care.</span><span style='font-size:14.0pt;line-height:
+
150%'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt;text-align:justify;line-height:150%'><span
+
style='font-size:14.0pt;line-height:150%;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
+
mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:text1;mso-font-kerning:
+
12.0pt'>(http://bioinformatics.psb.ugent.be/webtools/plantcare/html/)</span><span
+
style='font-size:14.0pt;line-height:150%'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt;text-align:justify;line-height:150%'><span
+
style='font-size:14.0pt;line-height:150%;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
+
mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:text1;mso-font-kerning:
+
12.0pt'>Functional validation of characterized MMV promoter via protoplast assay
+
in tobacco had done by expertise of the plant biotechnology lab, Department of
+
Botany D.E.I, <span class=SpellE>Dayalbagh</span>, <span class=GramE>Agra,</span>
+
Protocol had not been shared due to publication and patenting issues.<o:p></o:p></span></p>
+
 
+
<p align=center style='margin-top:0in;margin-right:0in;margin-bottom:0in;
+
margin-left:.5in;margin-bottom:.0001pt;text-align:center;text-indent:-.25in;
+
mso-list:l1 level1 lfo1'><![if !supportLists]><b><span style='font-size:20.0pt;
+
font-family:"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-fareast-font-family:
+
Calibri;color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'><span
+
style='mso-list:Ignore'>3.<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span></b><![endif]><b><span
+
style='font-size:20.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'>Protein
+
Characterization<o:p></o:p></span></b></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify;text-indent:-.25in;mso-list:l3 level1 lfo2'><![if !supportLists]><span
+
style='font-size:14.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;
+
mso-bidi-font-family:Symbol;color:#0D0D0D;mso-themecolor:text1;mso-themetint:
+
242'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span style='font-size:14.0pt;color:#0D0D0D;
+
mso-themecolor:text1;mso-themetint:242;mso-font-kerning:12.0pt'>Quantity
+
Estimation of Protein: For quantify the expressed protein Bradford Method is
+
used as given on the following link </span><span class=GramE><span
+
style='font-size:14.0pt;color:#0D0D0D;mso-themecolor:text1;mso-themetint:242'>-(</span></span><a
+
href="http://www.bio-protocol.org/e45"><span style='font-size:14.0pt;
+
color:#0D0D0D;mso-themecolor:text1;mso-themetint:242'>http://www.bio-protocol.org/e45</span></a><span
+
style='font-size:14.0pt;color:#0D0D0D;mso-themecolor:text1;mso-themetint:242;
+
mso-font-kerning:12.0pt'>).</span><span style='font-size:14.0pt;color:#0D0D0D;
+
mso-themecolor:text1;mso-themetint:242'><o:p></o:p></span></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify;text-indent:-.25in;mso-list:l3 level1 lfo2'><![if !supportLists]><span
+
style='font-size:14.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;
+
mso-bidi-font-family:Symbol;color:#0D0D0D;mso-themecolor:text1;mso-themetint:
+
242'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span style='font-size:14.0pt;color:#0D0D0D;
+
mso-themecolor:text1;mso-themetint:242;mso-font-kerning:12.0pt'>Quality
+
Estimation of protein:<span style='mso-spacerun:yes'>  </span>The quality of
+
expressed protein was done by SDS-PAGE.</span><span style='font-size:14.0pt;
+
color:#0D0D0D;mso-themecolor:text1;mso-themetint:242'><o:p></o:p></span></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify;text-indent:-.25in;mso-list:l3 level1 lfo2'><![if !supportLists]><span
+
class=MsoHyperlink><span style='font-size:14.0pt;font-family:Symbol;mso-fareast-font-family:
+
Symbol;mso-bidi-font-family:Symbol;color:#0D0D0D;mso-themecolor:text1;
+
mso-themetint:242;text-decoration:none;text-underline:none'><span
+
style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span></span><![endif]><span style='font-size:14.0pt;color:#0D0D0D;
+
mso-themecolor:text1;mso-themetint:242;mso-font-kerning:12.0pt'>Protocol for
+
the SD</span><span style='font-size:14.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
+
mso-bidi-theme-font:minor-bidi;color:#0D0D0D;mso-themecolor:text1;mso-themetint:
+
242;mso-font-kerning:12.0pt'>S-PAGE is followed as given on the following
+
web-link- (</span><a
+
href="http://www.assay-protocol.com/molecular-biology/electrophoresis/denaturing-page"><span
+
style='font-size:14.0pt;font-family:"Book Antiqua","serif";color:#0D0D0D;
+
mso-themecolor:text1;mso-themetint:242'>http://www.assay-protocol.com/molecular-biology/electrophoresis/denaturing-page</span></a><span
+
class=MsoHyperlink><span style='font-size:14.0pt;font-family:"Book Antiqua","serif";
+
color:#0D0D0D;mso-themecolor:text1;mso-themetint:242'>)</span></span><span
+
class=MsoHyperlink><span style='font-size:14.0pt;color:#0D0D0D;mso-themecolor:
+
text1;mso-themetint:242;text-decoration:none;text-underline:none'><o:p></o:p></span></span></p>
+
 
+
<p style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:.5in;
+
margin-bottom:.0001pt;text-align:justify'><span class=MsoHyperlink><span
+
style='font-size:14.0pt;color:#0D0D0D;mso-themecolor:text1;mso-themetint:242;
+
text-decoration:none;text-underline:none'><o:p>&nbsp;</o:p></span></span></p>
+
 
+
<p align=center style='margin-top:0in;margin-right:0in;margin-bottom:0in;
+
margin-left:.5in;margin-bottom:.0001pt;text-align:center;text-indent:-.25in;
+
mso-list:l1 level1 lfo1'><![if !supportLists]><b style='mso-bidi-font-weight:
+
normal'><span style='font-size:20.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;color:black;
+
mso-themecolor:text1'><span style='mso-list:Ignore'>4.<span style='font:7.0pt "Times New Roman"'>&nbsp;
+
</span></span></span></b><![endif]><b><span style='font-size:20.0pt;font-family:
+
"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>Immobilization of Cells Expressing our Protein
+
of Interest</span></b><span style='font-size:20.0pt'><o:p></o:p></span></p>
+
 
+
<p align=center style='margin:0in;margin-bottom:.0001pt;text-align:center'><b><span
+
style='font-size:20.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'><o:p>&nbsp;</o:p></span></b></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt;text-align:justify'><span
+
style='font-size:14.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'>Cells of<i> E. Coli</i>
+
Bl-21 expressing our desired protein are immobilized using the protocol given
+
by <span class=SpellE>Dhall</span> et al., 2012.</span><span style='font-size:
+
14.0pt'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt;text-align:justify'><span
+
style='font-size:10.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:black;mso-themecolor:text1;mso-font-kerning:12.0pt'>(http://sci-hub.cc/http://pubs.rsc.org/en/content/articlelanding/2012/ay/c2ay25960e/unauth#!divAbstract</span><span
+
style='font-size:10.0pt'><o:p></o:p></span></p>
+
 
+
<p align=center style='margin:0in;margin-bottom:.0001pt;text-align:center'><span
+
style='font-size:20.0pt'><o:p>&nbsp;</o:p></span></p>
+
 
+
<p align=center style='margin-top:0in;margin-right:0in;margin-bottom:0in;
+
margin-left:.5in;margin-bottom:.0001pt;text-align:center;text-indent:-.25in;
+
mso-list:l1 level1 lfo1'><![if !supportLists]><b style='mso-bidi-font-weight:
+
normal'><span style='font-size:20.0pt;font-family:"Calibri","sans-serif";
+
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;color:black;
+
mso-themecolor:text1'><span style='mso-list:Ignore'>5.<span style='font:7.0pt "Times New Roman"'>&nbsp;
+
</span></span></span></b><![endif]><b><span style='font-size:20.0pt;font-family:
+
"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>Efficacy Testing of Beads by treating with water
+
samples possessing heavy metals contamination</span></b><span style='font-size:
+
20.0pt'><o:p></o:p></span></p>
+
 
+
<p style='margin:0in;margin-bottom:.0001pt'><span style='font-size:14.0pt;
+
font-family:"Calibri","sans-serif";mso-ascii-theme-font:minor-latin;mso-bidi-font-family:
+
"Times New Roman";mso-bidi-theme-font:minor-bidi;color:black;mso-themecolor:
+
text1;mso-font-kerning:12.0pt'>Concentration of heavy metals in samples treated
+
with our fabricated bio-beads (BECO) are analyzed by ICP-<span class=GramE>OES<span
+
style='mso-spacerun:yes'>  </span>of</span> Agilent Technology</span><span
+
style='font-size:14.0pt;font-family:"Calibri","sans-serif";mso-ascii-theme-font:
+
minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;
+
color:white;mso-themecolor:light1;mso-font-kerning:12.0pt'>.</span><span
+
style='font-size:14.0pt'><o:p></o:p></span></p>
+
 
+
<p align=center style='margin:0in;margin-bottom:.0001pt;text-align:center'><span
+
style='font-size:20.0pt'><o:p>&nbsp;</o:p></span></p>
+
 
+
<p class=MsoNormal>References:<o:p></o:p></p>
+
 
+
<p class=MsoListParagraphCxSpFirst style='text-indent:-.25in;mso-list:l2 level1 lfo3'><![if !supportLists]><span
+
style='mso-bidi-font-family:Calibri;mso-bidi-theme-font:minor-latin'><span
+
style='mso-list:Ignore'>1.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span class=SpellE>Dhall</span> P, Kumar R and
+
Kumar A. 2012. BOD beads – a ready to use seeding material for estimation of
+
organic load of wastewater. Anal. Methods, 2012, 4, 4101. DOI:
+
10.1039/c2ay25960e<o:p></o:p></p>
+
 
+
<p class=MsoListParagraphCxSpMiddle style='text-indent:-.25in;mso-list:l2 level1 lfo3'><![if !supportLists]><span
+
style='mso-bidi-font-family:Calibri;mso-bidi-theme-font:minor-latin'><span
+
style='mso-list:Ignore'>2.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><![endif]><span style='font-size:10.5pt;line-height:115%;
+
font-family:"Open Sans","serif";color:black;background:white'>He, F. (2011).
+
Bradford Protein Assay.&nbsp;<em><span style='font-family:"Open Sans","serif";
+
mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi'>Bio-protocol</span></em>&nbsp;Bio101:
+
e45. DOI:&nbsp;</span><a href="https://doi.org/10.21769/BioProtoc.45"><span
+
style='font-size:10.5pt;line-height:115%;font-family:"Open Sans","serif";
+
color:#0094FF;background:white'>10.21769/BioProtoc.45</span></a><span
+
style='font-size:10.5pt;line-height:115%;font-family:"Open Sans","serif";
+
color:black;background:white'>.</span></p>
+
 
+
<p class=MsoListParagraphCxSpMiddle><span style='font-size:10.5pt;line-height:
+
115%;font-family:"Open Sans","serif";color:black;background:white'><o:p>&nbsp;</o:p></span></p>
+
 
+
<p class=MsoListParagraphCxSpMiddle><span style='font-size:10.5pt;line-height:
+
115%;font-family:"Open Sans","serif";color:black;background:white'><o:p>&nbsp;</o:p></span></p>
+
  
<p class=MsoListParagraphCxSpLast align=center style='text-align:center'><span
+
<p class=MsoNormal align=center style='text-align:center'><!--[if gte vml 1]><v:shapetype
style='mso-no-proof:yes'><!--[if gte vml 1]><v:shapetype id="_x0000_t75"
+
id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"
coordsize="21600,21600" o:spt="75" o:preferrelative="t" path="m@4@5l@4@11@9@11@9@5xe"
+
path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">
filled="f" stroked="f">
+
 
  <v:stroke joinstyle="miter"/>
 
  <v:stroke joinstyle="miter"/>
 
  <v:formulas>
 
  <v:formulas>
Line 1,122: Line 459:
 
  <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
 
  <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
 
  <o:lock v:ext="edit" aspectratio="t"/>
 
  <o:lock v:ext="edit" aspectratio="t"/>
</v:shapetype><v:shape id="Picture_x0020_1" o:spid="_x0000_i1025" type="#_x0000_t75"
+
</v:shapetype><v:shape id="Picture_x0020_8" o:spid="_x0000_s1026" type="#_x0000_t75"
  style='width:468pt;height:157.5pt;visibility:visible;mso-wrap-style:square'>
+
  alt="Description: C:\Users\aa\Desktop\2.jpg" style='position:absolute;left:0;
  <v:imagedata src="PROToCOLS_files/image001.emz" o:title=""/>
+
text-align:left;margin-left:13.5pt;margin-top:-8.2pt;width:521.75pt;height:292.2pt;
</v:shape><![endif]--><![if !vml]><img border=0 width=624 height=210
+
z-index:-251657216;visibility:visible;mso-wrap-style:square;
src="https://2017.igem.org/File:T-DEI-AGRA-INDIA-primage002.jpg"><![endif]></span></p>
+
mso-wrap-distance-left:9pt;mso-wrap-distance-top:0;mso-wrap-distance-right:9pt;
 +
mso-wrap-distance-bottom:0;mso-position-horizontal:absolute;
 +
mso-position-horizontal-relative:text;mso-position-vertical:absolute;
 +
mso-position-vertical-relative:text'>
 +
  <v:imagedata src="INTER_files/image001.jpg" o:title="2"/>
 +
<w:wrap type="tight"/>
 +
</v:shape><![endif]--><![if !vml]><img width=696 height=390
 +
src="INTER_files/image002.jpg" align=left hspace=12
 +
alt="Description: C:\Users\aa\Desktop\2.jpg" v:shapes="Picture_x0020_8"><![endif]><span
 +
style='font-size:56.0pt;line-height:115%'><o:p></o:p></span></p>
  
 
</div>
 
</div>

Revision as of 21:29, 1 November 2017

INTER-LAB STUDY

Description: C:\Users\aa\Desktop\2.jpg