Difference between revisions of "Team:KAIT JAPAN/lab-note"
| Line 2: | Line 2: | ||
<html> | <html> | ||
<main class="top-menu"> | <main class="top-menu"> | ||
| − | <h1>LAB NOTE</ | + | <h1><font color="#33B7FF">LAB NOTE</font></h1> |
| − | + | ||
<dl> | <dl> | ||
| − | <dt> | + | <dt>August 15th</dt> |
| − | <dd>Presentation at | + | <dd>Presentation at Promega for sponsorship of reagents.</dd> |
| − | <dt> | + | <dt>August 16th<dt/> |
| − | <dd>Transformed designated parts into DH5α<br>BBa_I20270,R0040,J364000,J364001,J364002,J364003,J364004,J364005</dd> | + | <dd>Transformed designated parts into DH5α<br>BBa_I20270, R0040, J364000 ,J364001, J364002, J364003, J364004, J364005</dd> |
| + | <dt>August 17th</dt> | ||
| + | <dd>Broth culture shows no growth</dd> | ||
| + | <dt>August 18th</dt> | ||
| + | <dd>Transformed designated parts into DH5α<br>BBa_I20270, R0040, J364000, J364001, J364002, J364003, J364004, J364005<br>Orders from IDT<br>・IL-10 receptor<br>・STAT1<br>・JAK<br>・GFAP promotor<br>・HlyB<br>・HlyD<br>・IL-12</dd> | ||
| + | <dt>August 19th</dt> | ||
| + | <dd>Colonies were formed on all plates<br>But only a small number of colonies were found on the plates I20270, J364004 and J364005.</dd> | ||
| + | <dt>August 20th</dt> | ||
| + | <dd>Open campus</dd> | ||
| + | <dt>August 21st</dt> | ||
| + | <dd>Transformed I20270, J364004 and J364005 into DH5α.</dd> | ||
| + | <dt>August 22nd</dt> | ||
| + | <dd>Precultured all 8 of the parts, then cultured them for 16 to 18 hours.</dd> | ||
| + | <dt>August 23rd</dt> | ||
| + | <dd>Carried out measurements for interLab study</dd> | ||
| + | <dt>August 24th</dt> | ||
| + | <dd>Organized the data measured<br>interLab study completed!</dd> | ||
| + | <dt>August 25th</dt> | ||
| + | <dd>Received gBlocks gene fragments ordered from IDT<br>Add-on orders from IDT<br>Collaboration with Vincent Lim from National Yang-Ming University></dd> | ||
| + | <dt>August 27th</dt> | ||
| + | <dd>PCR JAK5' and JAK3'</dd> | ||
| + | <dt>August 28th</dt> | ||
| + | <dd>PCR JAK5' and JAK3'</dd> | ||
| + | <dt>August 29th</dt> | ||
| + | <dd>PCR JAK5' and JAK3'</dd> | ||
| + | <dt>August 30th</dt> | ||
| + | <dd>1. Restriction digests and ligates JAK5' and JAK3'<br>2. PCR and gel purification of STAT1<br>3. PCR and gel purification of HlyD, HlyB and IL-12</dd> | ||
| + | <dt>August 31st</dt> | ||
| + | <dd>PCR of ligated JAK5'+3'(extension time 3:45)</dd> | ||
| + | <dt>September 1st</dt> | ||
| + | <dd>PCR ligated JAK5' and 3' (extension time 3:45) <br>diluted 100x, 1000x, 10000x restriction digests and gel purificated STAT1.<br>restriction digests and gel purification of HlyD, HlyB</dd> | ||
| + | <dt>September 3rd</dt> | ||
| + | <dd>Preculture pSB1C3<br>HlyB＋D ligation→colony PCR</dd> | ||
| + | <dt>September 4th</dt> | ||
| + | <dd>Minipreparation→electrophoresis<br>HlyB＋D→electrophoresis</dd> | ||
| + | <dt>September 5th</dt> | ||
| + | <dd>Minipreparation→gel purification restriction enzyme treatment (EcoRI, SpeI)<br>restriction digests PCR-ed HlyB＋D (EcoRI, XbaI)→gel purification</dd> | ||
| + | <dt>September 6th</dt> | ||
| + | <dd>pSB1C3 and HlyD+HlyB ligation⇨transformation<br>PCR:<br>1. GFP+HlyA<br>2. HlyA+GFP<br>3. HlyA+IL-12<br>4. IL-12+HlyA<br>5. diluted TolC</dd> | ||
| + | <dt>September 7th</dt> | ||
| + | <dd>Gel purification of pSB1C3<br>PCR:<br>1. GFP+HlyA<br>2. HlyA+GFP<br>3. HlyA+IL-12<br>4. IL-12+HlyA<br>5. TolC<br>→Gel purification | ||
| + | |||
| + | |||
| + | |||
| + | |||
| + | |||
Revision as of 07:54, 1 November 2017
LAB NOTE
- August 15th
- Presentation at Promega for sponsorship of reagents.
- August 16th
- Transformed designated parts into DH5α
BBa_I20270, R0040, J364000 ,J364001, J364002, J364003, J364004, J364005 - August 17th
- Broth culture shows no growth
- August 18th
- Transformed designated parts into DH5α
BBa_I20270, R0040, J364000, J364001, J364002, J364003, J364004, J364005
Orders from IDT
・IL-10 receptor
・STAT1
・JAK
・GFAP promotor
・HlyB
・HlyD
・IL-12 - August 19th
- Colonies were formed on all plates
But only a small number of colonies were found on the plates I20270, J364004 and J364005. - August 20th
- Open campus
- August 21st
- Transformed I20270, J364004 and J364005 into DH5α.
- August 22nd
- Precultured all 8 of the parts, then cultured them for 16 to 18 hours.
- August 23rd
- Carried out measurements for interLab study
- August 24th
- Organized the data measured
interLab study completed! - August 25th
- Received gBlocks gene fragments ordered from IDT
Add-on orders from IDT
Collaboration with Vincent Lim from National Yang-Ming University> - August 27th
- PCR JAK5' and JAK3'
- August 28th
- PCR JAK5' and JAK3'
- August 29th
- PCR JAK5' and JAK3'
- August 30th
- 1. Restriction digests and ligates JAK5' and JAK3'
2. PCR and gel purification of STAT1
3. PCR and gel purification of HlyD, HlyB and IL-12 - August 31st
- PCR of ligated JAK5'+3'(extension time 3:45)
- September 1st
- PCR ligated JAK5' and 3' (extension time 3:45)
diluted 100x, 1000x, 10000x restriction digests and gel purificated STAT1.
restriction digests and gel purification of HlyD, HlyB - September 3rd
- Preculture pSB1C3
HlyB＋D ligation→colony PCR - September 4th
- Minipreparation→electrophoresis
HlyB＋D→electrophoresis - September 5th
- Minipreparation→gel purification restriction enzyme treatment (EcoRI, SpeI)
restriction digests PCR-ed HlyB＋D (EcoRI, XbaI)→gel purification - September 6th
- pSB1C3 and HlyD+HlyB ligation⇨transformation
PCR:
1. GFP+HlyA
2. HlyA+GFP
3. HlyA+IL-12
4. IL-12+HlyA
5. diluted TolC - September 7th
- Gel purification of pSB1C3
PCR:
1. GFP+HlyA
2. HlyA+GFP
3. HlyA+IL-12
4. IL-12+HlyA
5. TolC
→Gel purification
Contact: kaitjapan@gmail.com Twitter:@KAIT_JAPAN
