IGEM:Peking/2007/Switch-Notebook/2007-7-6: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 53: Line 53:


30uL/2uL电泳看浓度
30uL/2uL电泳看浓度
[[Image:PKU Switch I13522-gfp(1-4)SD1&2.gif]]


===菌P(分区PCR)===
===菌P(分区PCR)===

Revision as of 18:57, 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.

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

  • MCS-pCC009(4uL/1uL), H20-pCC009自连

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)

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

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:Peking/2007/Switch-Notebook/2007-7-6&oldid=128182"