Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-8
... dNTPs ( if needed ) [ a - 32P ] dNTP H2O to 21.5 μl Chill the mixture to 0 ° C . 2.5 με 0.5 με 20 nmoles of each 16 pmoles For unlabeled dNTPs , add 1 μl of a 20 mM solution of each unlabeled dNTP ; usually , 20 nmoles of three unlabeled ...
... dNTPs ( if needed ) [ a - 32P ] dNTP H2O to 21.5 μl Chill the mixture to 0 ° C . 2.5 με 0.5 με 20 nmoles of each 16 pmoles For unlabeled dNTPs , add 1 μl of a 20 mM solution of each unlabeled dNTP ; usually , 20 nmoles of three unlabeled ...
Page 10-9
... dNTP . Most commercial suppliers sell [ a - 32P ] dNTPs in a concentrated aqueous solution , which can be added directly to the nick - translation reaction . The specific activities range from 400 to > 3000 Či / mmole . If [ a - 32P ] dNTPs ...
... dNTP . Most commercial suppliers sell [ a - 32P ] dNTPs in a concentrated aqueous solution , which can be added directly to the nick - translation reaction . The specific activities range from 400 to > 3000 Či / mmole . If [ a - 32P ] dNTPs ...
Page 10-20
... dNTPs . The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al . 1975 ) : dGTP 45 μM dCTP 200 μ dATP 12 μ dTTP 140 μ When the concentration of each of the four dNTPs in the reaction is greater than K , the ...
... dNTPs . The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al . 1975 ) : dGTP 45 μM dCTP 200 μ dATP 12 μ dTTP 140 μ When the concentration of each of the four dNTPs in the reaction is greater than K , the ...
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