Registry/Measurement kit/Notebook/2007-8-3
From OpenWetWare
(Difference between revisions)
| Line 1: | Line 1: | ||
| + | ==E0240== | ||
*Trying mutagenic PCR of E0240. | *Trying mutagenic PCR of E0240. | ||
| + | *Used 17 cycles. Don't know what # is optimal. Stratagene recommends 12, but people seem to use ~18. | ||
| + | **Too many cycles -> more junk DNA. | ||
| + | *Digesting with [[DpnI]]. Felix suggested digesting for >2h. | ||
| + | ==Transformations== | ||
*Later will try re-transforming into another set of competent Top10 cells. | *Later will try re-transforming into another set of competent Top10 cells. | ||
*Should make more MG1655. | *Should make more MG1655. | ||
| + | *Re-tried yesterday's transformations into Top10s. | ||
Revision as of 16:41, 3 August 2007
E0240
- Trying mutagenic PCR of E0240.
- Used 17 cycles. Don't know what # is optimal. Stratagene recommends 12, but people seem to use ~18.
- Too many cycles -> more junk DNA.
- Digesting with DpnI. Felix suggested digesting for >2h.
Transformations
- Later will try re-transforming into another set of competent Top10 cells.
- Should make more MG1655.
- Re-tried yesterday's transformations into Top10s.
