523-pCDNA3.1-Bcl-2

Cloning of pCDNA3.1-Bcl-2 (523)

Purpose: to clone pCDNA3.1-Bcl-2 by Gene Synthesis

Date: 2012-1-18 to 2012-1-23

Notebook: Xiaojing Wang

Page: 004163---004164

Clone ID: 523

Experimenters: Xiaojing Wang

Results

digest:

digest PCR product and vector with Hind111 +Xho1,run gel

the left band shows 523 insert, the right picture shows 335 vector (pCDNA3.1, 335)

gel purify, ligation, transformation

Colony Screening

Select 5 clones to mini-prep, digestd by Hind111 and Xho1. 523-1, -2, -3, -4 may be correct. Control is 523-1.

Select 523-1 to sequence.

Cloning of pCDNA3.1-Bcl-2 (523)

Purpose: to clone pCDNA3.1-Bcl-2 by Gene Synthesis

Date: 2012-3-13 to 2012-3-16

Notebook:

Page:

Clone ID: 523

Experimenters: yingying lu

Results

PCR:PCR using pDONR-Bcl-2, 1019 as template

oligos

Bcl-2-F C6 tgatcaAAGCTTGAATTCGCCGCCACCATG 10 uM
Bcl-2-2 CCGGTTATCGTACCCTGTTCTCCCAGCGTGCGCCATGGTGGCGGCGAATTCGTA 1 uM
Bcl-2-3 D8 GAACAGGGTACGATAACCGGGAGATAGTGATGAAGTACATCCATTATAAGCTGT 1 uM

and
Bcl-2-20 CACCCAGGGTGATGCAAGCTCCCACCAGGGCCAAACTGAGCAGAGTCTTCAGAG 1 uM
Bcl-2-21 CTTGCATCACCCTGGGTGCCTATCTGGGCCACAAGTGAGAATTCGCTCTCGAGG 1 uM
Bcl-2-R C7 tactcCTCGAGGAATTCTCACTTGTGGCCCAG 10 uM

	band1	band 2	band 3	band 4
	PCR product 3.12	335pCDNA3.1	PCR product 3.13	335pCDNA3.1
digest enzymes	Hind111 + Xho1		Hind111 + Xho1

clonies digest detect: Hind111 + Xho1

digest detect again:EcoR1

--Paul.xu 14:56, 16 March 2012 (EST)