Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 86
Page 12-18
... 4 ° C until the results of the immunological screening are available . If large areas of the top agarose stick to the nitrocellulose filters , chill the plates for 30 minutes at 4 ° C or 5 minutes at -20 ° C before peeling off the ...
... 4 ° C until the results of the immunological screening are available . If large areas of the top agarose stick to the nitrocellulose filters , chill the plates for 30 minutes at 4 ° C or 5 minutes at -20 ° C before peeling off the ...
Page 13-86
... 4 μl sonicated DNA 5 μl 32P - labeled DNA Chill to 0 ° C . Add 5 μl 10 % DMS and mix . Incubate 5 minutes at 20 ° C . Add 50 μl DMS stop solu- tion ( at 0 ° C ) and mix . Add 750 μl of ethanol ( -20 ° C ) and mix . Store for 5 minutes ...
... 4 μl sonicated DNA 5 μl 32P - labeled DNA Chill to 0 ° C . Add 5 μl 10 % DMS and mix . Incubate 5 minutes at 20 ° C . Add 50 μl DMS stop solu- tion ( at 0 ° C ) and mix . Add 750 μl of ethanol ( -20 ° C ) and mix . Store for 5 minutes ...
Page 15-42
... for at least 6 hours at 37 ° C . 4. Purify the DNA by extraction with phenol : chloroform . Separate the organic and aqueous phases by centrifugation at 12,000g for 3 minutes at 4 ° C in a microfuge , and transfer the aqueous phase to a ...
... for at least 6 hours at 37 ° C . 4. Purify the DNA by extraction with phenol : chloroform . Separate the organic and aqueous phases by centrifugation at 12,000g for 3 minutes at 4 ° C in a microfuge , and transfer the aqueous phase to a ...
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Common terms and phrases
agarose gel aliquots amino acid amplification bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA Biol buffer carried cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing deletions denatured digestion DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol ethidium bromide exonuclease formamide gel electrophoresis gene genomic DNA H₂O hybridization Incubate the reaction inserted Klenow fragment labeled ligation linkers method microfuge tube minutes at 4°C mismatched molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nucleotides oligonu oligonucleotide phagemid plaques plasmid plasmid DNA plates polymerase chain reaction pool primer probe protein purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples screening sequencing reactions single-stranded DNA solution specific activity Store strand of cDNA synthesis target DNA target fragment target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml