IGEM:Peking/2007/Switch-Notebook/2007-7-11

Experimenter: LCB, JLL

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!

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.



Check the plates of lacZa-pLX007
Pick whichever is better.

Send for sequencing
CD1-1, CD1-3, RD1-1

PCR
Various lacZa, pCC002-lacZa