Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-24
... ligase buffer 5 mM гATP 30 % w / v polyethylene glycol ( PEG 8000 ) ( optional - see note to step 1 ) H2O to 25 μl bacteriophage T4 DNA ligase for cohesive termini for blunt termini 2.5 με 2.5 με 5.0 με 1.0 Weiss unit 5.0 Weiss units ...
... ligase buffer 5 mM гATP 30 % w / v polyethylene glycol ( PEG 8000 ) ( optional - see note to step 1 ) H2O to 25 μl bacteriophage T4 DNA ligase for cohesive termini for blunt termini 2.5 με 2.5 με 5.0 με 1.0 Weiss unit 5.0 Weiss units ...
Page 15-11
... ligase therefore will ligate 50 % of the HindIII fragments of bacteriophage A ( 5 μg ) in 30 minutes at 16 ° C . Throughout this manual , bacteriophage T4 DNA ligase is given in Weiss units . 5. Digest the linkers with the appropriate ...
... ligase therefore will ligate 50 % of the HindIII fragments of bacteriophage A ( 5 μg ) in 30 minutes at 16 ° C . Throughout this manual , bacteriophage T4 DNA ligase is given in Weiss units . 5. Digest the linkers with the appropriate ...
Page 15-90
... 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 for 10 ...
... 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 for 10 ...
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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