Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 14-32
... original tenfold dilution of the control template ( step 2 ) . Do not reduce the concentration of unlabeled dCTP in the reaction mixture to increase the specific activity of the precursor pool . There is a danger that the concentration ...
... original tenfold dilution of the control template ( step 2 ) . Do not reduce the concentration of unlabeled dCTP in the reaction mixture to increase the specific activity of the precursor pool . There is a danger that the concentration ...
Page 15-36
... original target DNA are then separated from shorter fragments by gel electrophoresis and recloned into an appropriate plasmid vector . Because this fractionation step is at best accurate only to ± 5 nucleotide pairs , plasmids whose ...
... original target DNA are then separated from shorter fragments by gel electrophoresis and recloned into an appropriate plasmid vector . Because this fractionation step is at best accurate only to ± 5 nucleotide pairs , plasmids whose ...
Page 15-96
... original wild - type sequences . This type of mutagenesis is therefore best carried out using the Kunkel system ( see pages 15.74- 15.79 ) , which selects strongly against bacteriophages generated by replica- tion of the original wild ...
... original wild - type sequences . This type of mutagenesis is therefore best carried out using the Kunkel system ( see pages 15.74- 15.79 ) , which selects strongly against bacteriophages generated by replica- tion of the original wild ...
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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