PEDFR
digest pOTB7-PEDF-R ((ATCC # 10659852) with EcoR1 + Xho1. the size is about 600 bp, smaller than expected full-length clone 1.5 kb.
--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.
--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.
--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.
--Ztao 13:12, 10 February 2011 (EST)