IGEM:Peking/2007/Switch-Notebook/2007-7-6
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Experimenter: YT, YL, LCB, WMC
Contents
Precipate pLX007(SalI/XhoI), pCC009(SalI/XhoI) and lacZa(SalI/XhoI) with ethanol
2倍体积乙醇, 1/10体积NaAc.
回收于20uL, 离心30min.
Positive transformation of SDA-EGFP-pCC002(1,5) grown up
但提质粒用的是存下来的菌液, BamHI/XhoI(7uL/20uL)切检验, OK! 存质粒.
l to r: 1, 5, none, Marker
还有菌液在冰箱门上存着, 用时直接提.
mRFP-PCC002, SDY-ECFP-pCC002 plates do not grow, 昨天晚上连的体系转化
RD1-1-T, RD1-2-T transformation
Extract <bbpart>BBa_I13522</bbpart>(GFP)(1-4) from iGEM DNA Kit
Waiting for Mini-prep in the evening, labeled GFP-1,2,3,4.
Done.
Single digestion of pCC009(1-4), pLX007(4) with XhoI (2uL/20uL) to check
lacZa(SalI/XhoI)电泳看浓度, 10uL, Marker 10uL (稍大)
连接
- lacZa-pLX007(4uL/1uL), H2O-pLX007自连
l to r: pLX007^, pCC009(1-4), pLX007, Marker
- MCS-pCC009(4uL/1uL), H20-pCC009自连
l to r: lacZa(x/s), Marker
5pm-6pm: 4 degree, 6pm-9pm 16 degree.
KOD PCR SD1&2&3
结果如下, 切胶回收SD1&2较大片段, 30uL洗脱, Marker 10uL
l to r: SD1, SD2, SD3, M
连pMD18T (1hour, 16 degree), 转化
Miniprep of <bbpart>BBa_I13522</bbpart>(GFP)
OK! Stock in -20C fridge.
30uL/2uL电泳看浓度
l to r: I13522(GFP)(1-4), SD1, SD 2, Marker
菌P(分区PCR)
- RD1-1-pMD18T 1-12区
- RD1-2-pMD18T 1'-12'区
(每区3-5个不定, 以戳的洞为准)
Looking forward
检板子
- mRFP-pCC002
- SDY-ECFP-pCC002
- SD1-pMD18T
- SD2-pMD18T
- MCS-pCC009
- lacZa-pLX007
菌P结果电泳
- RD1-1-pMD18T
- RD1-2-pMD18T
如有对的提质粒酶剪切