Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 45
Denature the DNA by soaking the gel for 15 minutes in denaturation / transfer
solution ( 0 . 4 n NaOH , 1 m ... Transfer the denatured DNA from the gel to the
membrane by capillary action as described in steps 7 - 13 on page 9 . 39 . 5 .
Peel the ...
Denature the DNA by soaking the gel for 15 minutes in denaturation / transfer
solution ( 0 . 4 n NaOH , 1 m ... Transfer the denatured DNA from the gel to the
membrane by capillary action as described in steps 7 - 13 on page 9 . 39 . 5 .
Peel the ...
Page 14
Synthesis of Probes from Denatured Double - stranded DNA SYNTHESIS OF
RADIOLABELED PROBES BY PRIMER EXTENSION This method is used to
generate probes from denatured , closed circular DNA or denatured , linear ,
double ...
Synthesis of Probes from Denatured Double - stranded DNA SYNTHESIS OF
RADIOLABELED PROBES BY PRIMER EXTENSION This method is used to
generate probes from denatured , closed circular DNA or denatured , linear ,
double ...
Page 71
6 with acetic acid ) . Immediately mix the solution by vortexing briefly , and then
quickly add 50 ul of ethanol at 0°С . Mix again , and store the solution on ice for
15 minutes . 4 . Recover the denatured plasmid DNA by centrifugation at 12 ,
000g ...
6 with acetic acid ) . Immediately mix the solution by vortexing briefly , and then
quickly add 50 ul of ethanol at 0°С . Mix again , and store the solution on ice for
15 minutes . 4 . Recover the denatured plasmid DNA by centrifugation at 12 ,
000g ...
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Contents
TEST LIGATION 3 29 | 3 |
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 11 |
FRAGMENT OF E coli DNA POLYMERASE I 13 59 | 14 |
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Common terms and phrases
acid activity agarose aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies concentration constructed containing deletions denatured described determine digestion DNA polymerase dNTPs double-stranded efficiency electrophoresis et al ethanol expression Figure filters fragment gene genomic hybridization increase Incubate inserted interest isolated labeled length libraries ligation linear linkers method microfuge minutes mixture molecules mRNA mutagenesis mutagenic mutations Nucleic nucleotides obtained oligonucleotide original plaques plasmid plasmid DNA plate pool position possible prepared presence primer probe problem protein purified radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening sequence single single-stranded solution specific step Store strand synthesis target DNA target sequence template termini tion transfer transformants tube units usually vector volume