Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 36
Sometimes these colonies were pooled , amplified in liquid culture , and stored at
- 70°C ; more frequently , they were maintained on the surfaces of nitrocellulose
filters ( Hanahan and Meselson 1983 ) . In either case , the results were ...
Sometimes these colonies were pooled , amplified in liquid culture , and stored at
- 70°C ; more frequently , they were maintained on the surfaces of nitrocellulose
filters ( Hanahan and Meselson 1983 ) . In either case , the results were ...
Page 49
1984 ) is 5 % nonfat dried milk dissolved in water containing 0 . 02 % sodium
azide . It should be stored at 4°C and diluted 25 - fold into prehybridization buffer
before use . BLOTTO should not be used in combination with high concentrations
of ...
1984 ) is 5 % nonfat dried milk dissolved in water containing 0 . 02 % sodium
azide . It should be stored at 4°C and diluted 25 - fold into prehybridization buffer
before use . BLOTTO should not be used in combination with high concentrations
of ...
Page 48
10 % Ammonium persulfate Ammonium persulfate lg H , O to 10 ml The solution
may be stored at 4°C for several weeks . 6 % Acrylamidelurea top solution 40 %
acrylamide solution 75 ml 5 X TBE 50 ml urea ( ultrapure ) 230 g Adjust the ...
10 % Ammonium persulfate Ammonium persulfate lg H , O to 10 ml The solution
may be stored at 4°C for several weeks . 6 % Acrylamidelurea top solution 40 %
acrylamide solution 75 ml 5 X TBE 50 ml urea ( ultrapure ) 230 g Adjust the ...
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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