- Miniprep J04500 to get high volumes
- Midiprep J04500
- Digest J04500 /SP @ 2h @45 and KaiC /XP @ 2h @45
- CIP treat J04500 /SP
- Gel and purify both
- Vacufudge both to ~8uL total
Perry kindly volunteered to do the midiprep.
Sequencing of Sample "A", "E", "I", and "M" from yesterday
Used primer VR for each, expected ~500bp. Will get results tomorrow, submitted with Perry's stuff.
Digest 4500/SP and KaiC /XP
4 samples of each digested for 2h @ 45C. Run on gel; image pending.
Gel extraction and Post Digest CIP
Gel extracted the first two KaiC segments with each other and the each 4500/SP seperately; got around 30 ng/uL for the 4500/SP in buffer EB. CIP'd the 4 J4500 segments for 1h@37 using 1uL AP.
PCR Purification of 4 J4500 CIP products
PCR purified all 4 J4500 CIP products in one contaner; got 110ng/uL eluting in 30uL water, for around a 80% recovery.