IGEM:Peking/2007/Switch-Notebook/2007-7-6

Experimenter: YT, YL, LCB, WMC

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.

Extract BBa_I13522 (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 BBa_I13522 (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)

检板子

 * mRFP-pCC002
 * SDY-ECFP-pCC002
 * SD1-pMD18T
 * SD2-pMD18T
 * MCS-pCC009
 * lacZa-pLX007

菌P结果电泳
如有对的提质粒酶剪切
 * RD1-1-pMD18T
 * RD1-2-pMD18T