1714 pEC-iMMLV

Cloning of 1714 pEC-iMMLV

Purpose: Cloning of1714 pEC-iMMLV

Date: 2013-4-

Notebook:

Page:

Clone ID:1714

Experimenters: Shanyi

Results

cloning

Date:2013-3-

PCR use 1713 as template and primers

MMLV-EcoR1-Nde1-F + MMLV-E69K-R

MMLV-E69K-F + MMLV-E302R-R

MMLV-E302R-F + MMLV-N454K-R

MMLV-N454K-F + MMLV-D524N-R

MMLV-D524N-F + MMLV-Not1-Xho1-R

join 5 fragments by pcr using

MMLV-EcoR1-Nde1-F atcgac gaattc CAT ATG ACCCTAAATATAGAAGATGAGC

MMLV-Not1-Xho1-R (CTCCAGACACCTCTACCCTCCTC cat cac cat cac cat cac taa GCGGCCGC CTCGAG cgatca) tgatcgCTCGAGGCGGCCGCttagtgatggtgatggtgatgGAGGAGGGTAGAGGTGTCTGGAG

digest with  EcoR1 + Xho1

Colony Screening

Date: 2013

DIGEST WITH BamHI

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