Paul.xu: moved PEDFR Cloning to PEDFR

2011-05-06T23:25:49Z

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[[Image:POTB7-PEDF-R-EcoR1 Xho1.JPG]]

 

digest pOTB7-PEDF-R ((ATCC # 10659852) with EcoR1 + Xho1. the size is about 600 bp, smaller than expected full-length clone 1.5 kb. --[[User:Paul.xu|Paul.xu]] 16:43, 5 November 2010 (EST)

Clone PEDFR into pLVX-Plus2

PCR:

Digestion: digest PCR product with BamH1 + Xho1 for 1 hr at 37 oC.

Digest pLVX-Plus2 with BamH1 + Xho1

Gel purify,

ligate and transform STBL3

pick 5 clonies,

make miniprep.

Digest with BamH1+Xho1

 

From left to right:

1: marker;

2~6: PEDFR_pLVX_Plus2_1~5;

7: PEDFR-pLVX_Plus2_1 without cutting. 

Reaction: 1.5 ul 10 * buffer (fast), 3 ul plasmid, 0.5 ul BamHI, 0.5 ul XhoI, 9.5 ul ddH2O, total 15 ul. 37C, 1 hour. 

[[Image:PEDFR to pLVX-Plus.JPG]]

--[[User:Ztao|Ztao]] 09:19, 8 February 2011 (EST)

 

From left to right:

1: marker;

2~3: PEDFR_pLVX_Plus2_1, cut with BamHI or XhoI, respectively;

4~8: PEDFR-pLVX_Plus2_1~5: cut by BamHI and XbaI.

Reaction: 1.5 ul 10 * buffer (fast), 2 ul plasmid, 0.5 ul BamHI, 0.5 ul XbaI, 9.5 ul ddH2O, total 15 ul. 37C, 1 hour. 

[[Image:PEDFR to pLVX Plus 2-8-11.JPG|353x262px|PEDFR to pLVX Plus 2-8-11.JPG]] 

 --[[User:Ztao|Ztao]] 13:24, 8 February 2011 (EST)

 

From left to right:

1: marker;

2~6: PEDFR_pLVX_Plus2_1~5, cut with BamHI and XhoI.

Reaction: 1.5 ul 10 * buffer (fast), 2 ul plasmid, 0.5 ul BamHI, 0.5 ul XhoI, 9.5 ul ddH2O, total 15 ul. 37C, 1 hour. 

Run 1% agrose gel without dye. Put the gel to water containing dye after running for 30 min.

[[Image:PEDFR_to_pLVX_Plus_2-10-11.JPG|340x376px]] 
--[[User:Ztao|Ztao]] 13:12, 10 February 2011 (EST)

 

[[Category:Cloning]]

PEDFR - Revision history

Paul.xu: moved PEDFR Cloning to PEDFR