Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 64
Purify the DNA by extraction with phenol : chloroform and precipitation with 2
volumes of ice - cold ethanol for 15 minutes at 0°C . Recover the DNA by
centrifugation at 12 , 000g for 10 minutes at 4°C in a microfuge . 2 . Dissolve the
DNA in a ...
Purify the DNA by extraction with phenol : chloroform and precipitation with 2
volumes of ice - cold ethanol for 15 minutes at 0°C . Recover the DNA by
centrifugation at 12 , 000g for 10 minutes at 4°C in a microfuge . 2 . Dissolve the
DNA in a ...
Page 86
0 ) and mix . mix gently . Incubate 5 minutes at Incubate 15 minutes at Incubate 7
minutes at 20°C . 37°C . 20°C . Add 50 ul DMS stop solu - Add 240 ul hydrazine
Add 200 ul hydrazine tion ( at 0°C ) and mix . stop solution ( at 0°C ) stop solution
...
0 ) and mix . mix gently . Incubate 5 minutes at Incubate 15 minutes at Incubate 7
minutes at 20°C . 37°C . 20°C . Add 50 ul DMS stop solu - Add 240 ul hydrazine
Add 200 ul hydrazine tion ( at 0°C ) and mix . stop solution ( at 0°C ) stop solution
...
Page 24
5 minutes , 3 . 0 minutes , 4 . 5 minutes , etc . 3 . Add 36 ul of the appropriate
dilution of BAL 31 ( see step 6 , page 15 . 23 ) to the ... 5 - minute intervals ,
transfer 45 ul of the reaction mixture to the appropriately labeled microfuge tube .
Store the ...
5 minutes , 3 . 0 minutes , 4 . 5 minutes , etc . 3 . Add 36 ul of the appropriate
dilution of BAL 31 ( see step 6 , page 15 . 23 ) to the ... 5 - minute intervals ,
transfer 45 ul of the reaction mixture to the appropriately labeled microfuge tube .
Store the ...
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User Review - iayork - LibraryThingthe BIBLE of every biologist: So few and so much to say about this bible of Biology at the bench... You'll really find everything you want in it, including the composition of all the buffers and ... Read full review
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 |
Copyright | |
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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