Koeris/Notebook/2007-1-19: Difference between revisions
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#72 deg C - 10min | #72 deg C - 10min | ||
#4 deg C - indefinite | #4 deg C - indefinite | ||
==Gel image== | |||
==Restriction digest== | |||
Digest the PCR amplicons with KpnI, Hind III - double digest | |||
Revision as of 08:24, 19 January 2007
Re-cloning of [His]6
Due to lack of material, I could not recover the A plasmid and have to re-clone it.
PCR set-up
- 100 ul total reaction volume
- 1ul template DNA @ ~40ng/ul
- 200pmol primer mix
- 97ul Taq SuperMix
Pipetting scheme
Labels in the respective field codes
| Primer | Tube Label |
|---|---|
| A his6 2F&R 200pmol | A1 |
| A his6 2F&R 200pmol | A2 |
Thermal profile
- 92 deg C - 5min
- 92 deg C - 30s
- 50 deg C - 45s
- 72 deg C - 120s
- Cycle back to 2. 29X
- 72 deg C - 10min
- 4 deg C - indefinite
Gel image
Restriction digest
Digest the PCR amplicons with KpnI, Hind III - double digest
