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[PCR] DNA-se I FOOTPRINTING 日期：2005-07-18 00:00:00 点击：335 好评：0
Friday, November 21, 2003 Description This technique exploits the fact that a protein bound to a specific DNA sequence will interfere with the digestion of that region by the endonuclease DNAaseI. In summary, an end-labelled DNA probe is i...
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[PCR] Nested Deletions using exonuclease-III and mung bean nucleas 日期：2005-07-18 00:00:00 点击：203 好评：0
Friday, November 21, 2003 Description Nested Deletions using exonuclease-III and mung bean nuclease Procedure ExoIII has a 3' to 5' exonuclease activity which must initiate on blunt ended DNA or DNA with a 5' overhang. 1. A nested series o...
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[PCR] Nested Deletions using exonuclease-III and mung bean nucleas 日期：2005-07-18 00:00:00 点击：307 好评：0
Friday, November 21, 2003 Description Nested Deletions using exonuclease-III and mung bean nuclease Procedure ExoIII has a 3' to 5' exonuclease activity which must initiate on blunt ended DNA or DNA with a 5' overhang. 1. A nested series o...
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[PCR] In vitro mutagenesis using Altered Sites 日期：2005-07-18 00:00:00 点击：293 好评：0
Friday, November 21, 2003 Description In vitro mutagenesis using Altered Sites Procedure A. Isolation of Single Stranded DNA 1. Transform plasmid to be mutagenised into JM109. JM109 is endA1, recA1, gyrA96, thi, hsdR17(rk-, mk+), relA1, su...
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[PCR] Ligation 日期：2005-07-18 00:00:00 点击：818 好评：0
Tuesday, November 18, 2003 Description Appropriate vector DNA is digested with the relevent restriction endonucleases and the 5'-terminal phosphates removed by CIP. The DNA fragment to be cloned is typically prepared by endonuclease digest...
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[PCR] 3RACE PCR 日期：2005-07-18 00:00:00 点击：465 好评：0
Tuesday, November 18, 2003 Description 3' Rapid Amplification of cDNA Ends (RACE) PCR This technique is used to obtain the 3'end of a cDNA, it requires some sequence information internal to the mRNA under study. The sequence information ob...
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[PCR] Disruption by Fusion PCR 日期：2005-07-18 00:00:00 点击：289 好评：0
Tuesday, November 18, 2003 Description Disruption by Fusion PCR Procedure 1) In separate PCR reactions, amplify the 5' and 3' ends of the gene of interest with primers about 200 bases apart. Primer 2 should begin with 24 nts complementary...
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[PCR] UV Mutagenesis 日期：2005-07-18 00:00:00 点击：328 好评：0
Tuesday, November 18, 2003 Description UV Mutagenesis Procedure 1) Grow cells 2-3 days to stationary phase 2) Wash cells 2x with water 3) Count cells (vortex well to separate cells as much as possible) 4) Resuspend to 2 x 108 cells/ml 5) T...
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[PCR] Vectorette PCR of Yeast DNA 日期：2005-07-18 00:00:00 点击：248 好评：0
Tuesday, November 18, 2003 Description Vectorette PCR of Yeast DNA Procedure 1) Cut 1-3 g of clean DNA overnight with 8-10U of blunt cutting enzyme in 20l Most problems come from dirty, uncut DNA. Phenol/glass bead/RNase prepared DNA works...
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[PCR] Yeast Colony PCR 日期：2005-07-18 00:00:00 点击：312 好评：0
Tuesday, November 18, 2003 Description Yeast Colony PCR Procedure PCR mixture Combine reaction mix on ice: 2.5 μl 10X Colony PCR Buffer 1.5 μl 25 mM MgCl2 0.5 μl 10 mM dNTP's 10 pmols of each primer 1.25 μl 20% Triton X-100 0.25 μl Ta...