1193 pCDNA3.2-Fc

pCDNA3.2-Fc (1193)

Date: 2012-05-26

Notebook:  4480

Page:  4481

Clone ID: 1193

Experimenters: Paul

Cloning

PCR Fc domain from pFUSE-hIgG1-Fc2-IAB 1101 using these oligos:
ssFc-BamH1-F1 + 10% ss-Fc-F5 + Fc-Kpn1-R4, template is pFUSE-hIgG1-Fc2-IAB, 1101
ssFc-BamH1-F1
accgtcGGATCCGCCACCATGGGAGTCAAAGTTCTGTTTGCCCTGATCTGCATCGCTGTG
GGATCCGCCACCATGGGAGTCAAAGTTCTGTTTGCCCTGATCTGCATCGCTGTG GCC GAG GCC AAG CCC ACC
ss-Fc-F5
CCCTGATCTGCATCGCTGTGGCCGAGGCCAAGCCCACCgacaaaactcacacatgcccac
Fc-Kpn1-R4
(ctctccctgtctccgggtaaaGGCAGCGGCGAGAACCTGTACTTCCAGGGCggtaccATCGAC)
tactagggtaccGCCCTGGAAGTACAGGTTCTCGCCGCTGCCtttacccggagacagggagag

Digest with BamH1 + Kpn1, clone into pCDNA3.2, 1192

5/27/2012

digest PCR product and vector with BamH1 + Kpn1

1	2	3	4
1192 pCDNA3.2	1198 ssRFP-Fc-(3c)	1193 Fc	1199 ssRFP-Fc-(tev)

gel purified, and ligate, transform DH5alpha.

Colony Screening by Restriction Digestion

5-29-2012

clone 2,4,6 may be right, neeed to sequence