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Team:TU Munich/Notebook/Labjournal
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Revision as of 16:02, 23 June 2012
Contents |
Saturday, 23th June
Quick Change mutagenis to remove NgoMIV from pYES2
Investigator: Ingmar, Volker Aim of the experiment: Generation of an RFC 25 compatible version of the pYes2 Vector.
PCR reaction batch
| volume | reagent |
| 2.5 µl | 10x Pfu Ultra II buffer |
| 4 µl | Plasmid P7 pYes2_RFC25 MCS template |
| 0.5 µl | 1:10 dilution of O38 (10 pmol/µL) |
| 0.5 µl | 1:10 dilution of O39 ((10 pmol/µL) |
| 17 µl | ddH2O |
| 0.5 µl | dNTP mix |
| 0.5 µl | Pfu Turbo DNA polymerase (2.5 U / µl) |
PCR cycling parameters
| Segment | Cycles | Temperature | Time |
| 1 | 1 | 95 °C | 30 sec |
| 2 | 15 | 95°C | 30 sec |
| 55°C | 1 min | ||
| 68°C | 6 min |
