Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-56
... T4 DNA ligase Required only if self - priming of second - strand cDNA synthesis in un- acceptably high ( see note to step 5g on page 8.65 ) . Preparations of actinomycin D supplied by pharmaceutical manufacturers for therapeutic uses ...
... T4 DNA ligase Required only if self - priming of second - strand cDNA synthesis in un- acceptably high ( see note to step 5g on page 8.65 ) . Preparations of actinomycin D supplied by pharmaceutical manufacturers for therapeutic uses ...
Page 15-11
... T4 DNA ligase . Most manufacturers ( apart from New England Biolabs ) now calibrate the enzyme in Weiss units ( Weiss et al . 1968 ) . One Weiss unit is the amount of enzyme that catalyzes the exchange of 1 nmole of 32P from ...
... T4 DNA ligase . Most manufacturers ( apart from New England Biolabs ) now calibrate the enzyme in Weiss units ( Weiss et al . 1968 ) . One Weiss unit is the amount of enzyme that catalyzes the exchange of 1 nmole of 32P from ...
Page 15-90
... T4 DNA ligase and continue incubation for a further 1 hour at 16 ° C . 10x Ligase buffer 0.2 м Tris Cl ( pH 7.6 ) 50 mM MgCl2 50 mм dithiothreitol For definition of Weiss unit , see page 15.30 . iii . Heat the ligation mixture to 68 ° C ...
... T4 DNA ligase and continue incubation for a further 1 hour at 16 ° C . 10x Ligase buffer 0.2 м Tris Cl ( pH 7.6 ) 50 mM MgCl2 50 mм dithiothreitol For definition of Weiss unit , see page 15.30 . iii . Heat the ligation mixture to 68 ° C ...
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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