IGEM:Peking/2007/Switch-Notebook/2007-7-6: Difference between revisions
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===RD1-1-T, RD1-2-T转化=== | ===RD1-1-T, RD1-2-T转化=== | ||
===提取<bbpart>BBa_I13522</bbpart>(1-4)=== | ===提取<bbpart>BBa_I13522</bbpart>(GFP)(1-4)=== | ||
等晚上提质粒. 试管上标记为GFP-1,2,3,4. | 等晚上提质粒. 试管上标记为GFP-1,2,3,4. | ||
| Line 34: | Line 34: | ||
* MCS-pCC009(4uL/1uL), H20-pCC009自连 | * MCS-pCC009(4uL/1uL), H20-pCC009自连 | ||
[[Image:PKU_Switch_07-7- | [[Image:PKU_Switch_07-7-6_lazZa_xs.gif]] | ||
5pm-6pm: 4 degree, 6pm-9pm 16 degree. | 5pm-6pm: 4 degree, 6pm-9pm 16 degree. | ||
| Line 48: | Line 48: | ||
连pMD18T (1hour, 16 degree), 转化 | 连pMD18T (1hour, 16 degree), 转化 | ||
===<bbpart>BBa_I13522</bbpart>(GFP)提质粒=== | |||
OK! 存好. | |||
30uL/2uL电泳看浓度 | |||
===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) | |||
Revision as of 08:09, 6 July 2007
Experimenter: YT, YL, LCB, WMC
Contents
乙醇沉淀pLX007(SalI/XhoI), pCC009(SalI/XhoI) and lacZa(SalI/XhoI)
2倍体积乙醇, 1/10体积NaAc.
回收于20uL, 离心30min.
SDA-EGFP-pCC002(No.1,5)阳转长起来了
但提质粒用的是存下来的菌液, BamHI/XhoI(7uL/20uL)切检验, OK! 存质粒.
.gif)
还有菌液在冰箱门上存着, 用时直接提.
mRFP-PCC002, SDY-ECFP-pCC002的板子没有长, 昨天晚上连的体系转化
RD1-1-T, RD1-2-T转化
提取<bbpart>BBa_I13522</bbpart>(GFP)(1-4)
等晚上提质粒. 试管上标记为GFP-1,2,3,4.
已提好.
pCC009(1-4), pLX007(4) XhoI单切检验(2uL/20uL)
lacZa(SalI/XhoI)电泳看浓度, 10uL, Marker 10uL (稍大)
连接
- lacZa-pLX007(4uL/1uL), H2O-pLX007自连
pCC009(1-4)XhoI.jpg)
- MCS-pCC009(4uL/1uL), H20-pCC009自连
File:PKU Switch 07-7-6 lazZa xs.gif
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
- run time: 15 min
- run time: 30 min
连pMD18T (1hour, 16 degree), 转化
<bbpart>BBa_I13522</bbpart>(GFP)提质粒
OK! 存好.
30uL/2uL电泳看浓度
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)
