Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-24
... ligase by heating the mixture to 68 ° C for 15 minutes . 10 × Bacteriophage T4 DNA ligase buffer . 200 mM Tris Cl ( pH 7.6 ) 50 mM MgCl2 50 mm dithiothreitol 500 μg / ml bovine serum albumin ( Fraction V ; Sigma ) ( optional ) This ...
... ligase by heating the mixture to 68 ° C for 15 minutes . 10 × Bacteriophage T4 DNA ligase buffer . 200 mM Tris Cl ( pH 7.6 ) 50 mM MgCl2 50 mm dithiothreitol 500 μg / ml bovine serum albumin ( Fraction V ; Sigma ) ( optional ) This ...
Page 15-30
... ligase therefore will ligate 50 % of the HindIII fragments of bacteriophage λ ( 5 μg ) in 30 minutes at 16 ° C . Throughout this manual , bac- teriophage T4 DNA ligase is given in Weiss units . 14. Heat the ligation reaction to 68 ° C ...
... ligase therefore will ligate 50 % of the HindIII fragments of bacteriophage λ ( 5 μg ) in 30 minutes at 16 ° C . Throughout this manual , bac- teriophage T4 DNA ligase is given in Weiss units . 14. Heat the ligation reaction to 68 ° C ...
Page 15-90
... ligase and continue incubation for a further 1 hour at 16 ° C . 10x Ligase buffer · 0.2 M 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 for ...
... ligase and continue incubation for a further 1 hour at 16 ° C . 10x Ligase buffer · 0.2 M 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 for ...
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Common terms and phrases
acrylamide agarose gel aliquots amplification antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter chromatography cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing ddNTPs deletions denatured described digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers method MgCl2 microfuge tube minutes at 4°C molecules mRNA mutants Natl nitrocellulose nitrocellulose filters nucleic acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing reactions solution specific activity step stored strand of cDNA synthesis target DNA target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml