Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 49
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 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 ...
... DNA 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 ...
Page 15-90
... DNA 10 ligase buffer H2O to 9 μl 0.5 με 2 μ . 1 με Add 2 Weiss units of bacteriophage T4 DNA ligase and 10 mм ATP to a final concentration of 1 mм . Incubate the mixture for 12-16 hours at 4 ° C , and then add another aliquot ( 2 Weiss ...
... DNA 10 ligase buffer H2O to 9 μl 0.5 με 2 μ . 1 με Add 2 Weiss units of bacteriophage T4 DNA ligase and 10 mм ATP to a final concentration of 1 mм . Incubate the mixture for 12-16 hours at 4 ° C , and then add another aliquot ( 2 Weiss ...
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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