332-pLenti-RFP-LoxP-GFP

Cloning of pLenti-RFP-LoxP-GFP (332)

Purpose: to clone pLenti-RFP-LoxP-GFP by PCR

Date: 2011-9-20 to 2011-9-

Notebook: Xiaojing Wang

Page: 004095---004096

Clone ID: 332

Experimenters: Xiaojing Wang

Results

TagGFP was amplified by gene synthesis.

The right band shows PCR product of LoxP-TagGFP, TagGFP as template

LoxP-TagGFP-EcoR1-F C4 atcga gaattc ATAACTTCGTATA ATG TAT GC T ATA CGA AGT TAT AGC GGG GGC GAG GAG CTG

TagGFP-R1 XhoI (CACGGCATGGACGAGCTGTACAGGTAA ctcgag atcga) tcgatctcgagTTACCTGTACAGCTCGTCCATGCCGTG

Digest result:

digest PCR product and pLenti-mKate2 vector with 0.5ul EcoR1+ 0.8ul Xho1 at 37 for 1 hour , run gel

gel purify, ligate, transform STBL3  (Amp)

Colony Screening