Difference between revisions of "Team:WHU-China/Experiments"
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<h1 id="Materials">Materials</h1> | <h1 id="Materials">Materials</h1> | ||
<table border="1" align="center"> | <table border="1" align="center"> | ||
| − | <tr bgcolor="#89B9F8"><td style="text-align:center;vertical-align:middle;">Solutions</td><td | + | <tr bgcolor="#89B9F8"><td style="text-align:center;vertical-align:middle;">Solutions</td><td style="text-align:center;vertical-align:middle;">Ingredients</td><td style="text-align:center;vertical-align:middle;">Sterilization</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">2x AB3(Antibiotic Medium No.3)</td><td style="text-align:center;vertical-align:middle;">35g/L AB3(DIFCO) in dH2O</td><td style="text-align:center;vertical-align:middle;">Autoclave at 121℃ for 20 min</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">2×SMM </td><td style="text-align:center;vertical-align:middle;">40mM Maleic acid, 80mM NaOH, 40mM MgCl2•6H2O, 1M Sucrose in dH2O</td><td style="text-align:center;vertical-align:middle;"> Filtration</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">PEG-P</td><td style="text-align:center;vertical-align:middle;">400g/L PEG-6000 in 1x SMM</td><td style="text-align:center;vertical-align:middle;">Autoclave at 121℃ for 11 min</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">SMMP</td><td style="text-align:center;vertical-align:middle;">2x AB3 : 2×SMM= 1:1 </td><td style="text-align:center;vertical-align:middle;">Separately sterilized</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">Solution A</td><td style="text-align:center;vertical-align:middle;">206g/L Sucrose, 13g/L MOPS, 1.2g/L NaOH in dH2O.Adjust to pH 7.3 with NaOH </td><td style="text-align:center;vertical-align:middle;">Filtration</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">Solution B</td><td style="text-align:center;vertical-align:middle;">14.04g/L Agar, 0.70g/L Casamino acids, 35.09g/L Yeast Extract in dH2O</td><td style="text-align:center;vertical-align:middle;">Autoclave at 121℃ for 15 min</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">8x CR5 salts</td><td style="text-align:center;vertical-align:middle;">2g/L K2SO4, 80g/L MgCl2•6H2O, 0.4g/L KH2PO4, 17.6g/L CaCl2 in dH2O</td><td style="text-align:center;vertical-align:middle;">Autoclave at 121℃ for 15 min</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">12% Proline 120g/L</td><td style="text-align:center;vertical-align:middle;">L-proline in dH2O</td><td style="text-align:center;vertical-align:middle;">Filtration</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">20% Glucose</td><td style="text-align:center;vertical-align:middle;">200g/L Glucose in dH2O</td><td style="text-align:center;vertical-align:middle;">Filtration</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">CR5-top agar(2.5ml) </td><td style="text-align:center;vertical-align:middle;">1.25ml Solution A, 288μl CR5 salts, 125μl 12% Proline, 125μl 20% Glucose, 713μl Solution B(42℃)</td><td style="text-align:center;vertical-align:middle;">Separately sterilized and mix in order</td></tr> |
| − | <tr><td | + | <tr><td style="text-align:center;vertical-align:middle;">Lysis buffer</td><td style="text-align:center;vertical-align:middle;">100mM Na3PO4, 5mg/ml Lysozyme. Adjust to pH 6.3 with H3PO4. 1μl 1 M MgSO4 solution and 2 μl HS-Nuclease* (250 U/μl, cat.# GENUC10700-01, final concentration of 500 U/ml)</td><td style="text-align:center;vertical-align:middle;">Filtration</td></tr> |
</table> | </table> | ||
Revision as of 00:55, 30 October 2017
Materials
| Solutions | Ingredients | Sterilization |
| 2x AB3(Antibiotic Medium No.3) | 35g/L AB3(DIFCO) in dH2O | Autoclave at 121℃ for 20 min |
| 2×SMM | 40mM Maleic acid, 80mM NaOH, 40mM MgCl2•6H2O, 1M Sucrose in dH2O | Filtration |
| PEG-P | 400g/L PEG-6000 in 1x SMM | Autoclave at 121℃ for 11 min |
| SMMP | 2x AB3 : 2×SMM= 1:1 | Separately sterilized |
| Solution A | 206g/L Sucrose, 13g/L MOPS, 1.2g/L NaOH in dH2O.Adjust to pH 7.3 with NaOH | Filtration |
| Solution B | 14.04g/L Agar, 0.70g/L Casamino acids, 35.09g/L Yeast Extract in dH2O | Autoclave at 121℃ for 15 min |
| 8x CR5 salts | 2g/L K2SO4, 80g/L MgCl2•6H2O, 0.4g/L KH2PO4, 17.6g/L CaCl2 in dH2O | Autoclave at 121℃ for 15 min |
| 12% Proline 120g/L | L-proline in dH2O | Filtration |
| 20% Glucose | 200g/L Glucose in dH2O | Filtration |
| CR5-top agar(2.5ml) | 1.25ml Solution A, 288μl CR5 salts, 125μl 12% Proline, 125μl 20% Glucose, 713μl Solution B(42℃) | Separately sterilized and mix in order |
| Lysis buffer | 100mM Na3PO4, 5mg/ml Lysozyme. Adjust to pH 6.3 with H3PO4. 1μl 1 M MgSO4 solution and 2 μl HS-Nuclease* (250 U/μl, cat.# GENUC10700-01, final concentration of 500 U/ml) | Filtration |
(Other basic molecular biological materials are not listed here)
