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Team:BYU Provo/Notebook/Cholera - Enzymes/Protocols
From 2013.igem.org
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*0.5 ul Phusion Polymerase | *0.5 ul Phusion Polymerase | ||
| - | Set the PCR machine to run the following cycle | + | Set the PCR machine to run the following cycle with the middle three being repeated 35 times |
*98°C for 02:00 | *98°C for 02:00 | ||
| - | |||
*98°C for 00:30 | *98°C for 00:30 | ||
*65°C for 00:30 | *65°C for 00:30 | ||
*72°C for 04:00 | *72°C for 04:00 | ||
| - | |||
*72°C for 8:00 | *72°C for 8:00 | ||
Hold at 4°C | Hold at 4°C | ||
| Line 94: | Line 92: | ||
*0.5 ul ''TAQ'' Polymerase | *0.5 ul ''TAQ'' Polymerase | ||
| - | 95 for 2 | + | *95 for 2:00 |
| - | 95 for | + | *95 for 0:30 |
| - | 50 for | + | *50 for 0:30 |
| - | 72 for 1 | + | *72 for 1:00 |
| - | 72 for 1 | + | *72 for 1:00 |
<br> | <br> | ||
===Any deviations from these protocols are listed in the notebook when they were used=== | ===Any deviations from these protocols are listed in the notebook when they were used=== | ||
Revision as of 21:47, 27 September 2013
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Cholera-Enzyme Protocols
Salt LB recipe
To 950 ml distilled H2O add
Adjust pH to 7.8
Fill to 1000 ml and autoclave
Phusion PCR (50ul reaction)
For each sample add:
Set the PCR machine to run the following cycle with the middle three being repeated 35 times
Hold at 4°C
Taq polymerase PCR (50ul reaction)
For each sample add:
===Any deviations from these protocols are listed in the notebook when they were used=== |
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