IGEM:Peking/2007/Switch-Notebook/2007-7-8
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Experimenter: LXL, NM, CCY
Contents
Send for sequencing RD1-1,2, SD1,2
Positive transformation of pLX007, 挑菌摇2个
pMDB与pMDB0的效率差别不大
(当然还是少了一点的)
看来感受态放在-40C还是可以的, 上次应该是运输的问题, 下次可以用liquid nitrogen直接带来
Colony PCR test of SD1,2, lacZa-pMD18T
- primers: M13F/R
- control: positive transformation of pMDB
l to r: Marker, Negative control, SD1(1-12), Negative control, SD2(1-10)
l to r: Marker, SD2(11-12), Negative control, lacZa(1-11)
For sequencing tomorrow: SD1(1,4,5), SD2(1,2,3)
For miniprep tonight: lacZa(1,2,3)
ECFP^^失活, precipitate with enthanol
- precipitate at 9:30am
- collect at 2pm
Miniprep of ECFP1-5, mRFP1-20, 酶切检测
l to r: ECFP(1-5), mRFP(1-19), Marker
- ECFP is not ligated.
- mRFP(2,4,14,18) are correct.
SDE-EGFP-pMD18T酶切
l to r: Marker, mRFP20, SDE-EGFP-pMD18T^^
What’s wrong?
Re-try:
l to r: Marker, SDE-EGFP-pMD18T, ECFP^^
OK!
KOD PCR of SDD, SDD’_EGFP
- Primers: SDD_EGFP_BamHI_F, SDD’_EGFP_BamHI_F (newly ordered) + GFP-ClaI-R
- Template: SDD_EGFP_pMD18T
l to r: SDD, SDD', none, Marker, SDE-EGFP-pMD18T^^
切胶回收, 双切过夜 (BamHI & Cla I)
Result of pCC002-ECFP
Failed. pCC002自连太多, 于是失败. pCC002重切过夜.
ECFP^^回收, 连接, 过夜
- ECFP, pCC002, ClaI/NotI
pCC002^^
Miniprep of pLX007, lacZa (xs)-pMD18T
No SalI.
