Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 47
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 ... Ligation of the Target DNA 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 ... Ligation of the Target DNA 13 24.
Page 15-30
... 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 ... DNA LIGASES, KINASES, AND PHOSPHATASES 5 61 Bacteriophage T4 DNA Ligase 5 62.
... 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 ... DNA LIGASES, KINASES, AND PHOSPHATASES 5 61 Bacteriophage T4 DNA Ligase 5 62.
Page 15-90
... DNA ligase and continue incubation for a further 1 hour at 16 ° C . 10 x 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 ...
... DNA ligase and continue incubation for a further 1 hour at 16 ° C . 10 x 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 ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
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acrylamide agarose gel aliquots amplification annealing antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g cleavage cleaved cleotide coli DNA polymerase concentration containing ddNTPs deletions denatured digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers Maxam-Gilbert method MgCl2 microfuge tube minutes at 4°C molecules mRNA Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid piperidine plaques plasmid plasmid DNA plate polymerase chain reaction precipitation prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing gel sequencing reactions sonicated specific activity stored strand of cDNA synthesis T4 DNA polymerase target DNA target sequence template DNA termini transcription transfer Tris Cl pH vectors µg/ml