IGEM:Peking/2007/Switch-Notebook/2007-7-11
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Experimenter: LCB, JLL
Contents
Precipitate with ethanol: pLX007 (X/S)
- Time: 8:30-12:30
Temperature PCR of SD2 (X/S)
- Template: SD2-pMD18T
- Primers: lacO2_XhoI_XbaI_F, lacO2_SalI_SpeI_R
(Woooo.. I forgot to take the picture..)
Failed.
Colony PCR test of SDD’-EGFP-pCC002, RD1, CD1, SS1-3, SD3-pMD18T
l to r: Negative control, SS1(1-10), SD3(13-18), SS3(25-30), RD1, Marker
l to r: SS3(31-32), SS2(37-44), SDD'-EGFP-pCC002(1-12), none, Negative control for SS2, Marker
SDD'-EGFP-pCC002(13-35), Negative Control, Marker
Send for sequencing: bla…bla…bla…
收ECFP-pCC002新连的板子
自连&连接长得差不多!
Ligation of pLX007 (X/S) and lacZa
Ordered primers arrived
- SDA-H, cro, Y_lacZa_NotI_F
PCR test of CD1(1-6), RD1(1-6)-pMD18T
Photo omitted. The result is not good. We need to repeat this.
PCR of various SD_lacZa
Need to be repeated.
Colony PCR test of pLX007-lacZa
- testF&R
Failed..
Ligation and transformation
- lacZa and pLX007, 4:1 or 4.7:0.3
- positive transformation of pCC002-lacZa into JM109 and Math-T1
Sequencing result received
SD1-3, SD2-1, SD2-2 are right.
Miniprep and digestion test with BamHI.
Looking forward
Precipitate pLX007 (S/X)
Check the plates of lacZa-pLX007
Pick whichever is better.
收pCC002-lacZa板子到4C
Send for sequencing
CD1-1, CD1-3, RD1-1
PCR
Various lacZa, pCC002-lacZa
