Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-8
5 ug 20 nmoles of each 16 pmoles For unlabeled dNTPs , add 1 ul of a 20 mM
solution of each unlabeled dNTP ; usually , 20 nmoles of three unlabeled dNTPs
are added to the nick - translation mixture , e . g . , DTTP , DATP , and dGTP .
5 ug 20 nmoles of each 16 pmoles For unlabeled dNTPs , add 1 ul of a 20 mM
solution of each unlabeled dNTP ; usually , 20 nmoles of three unlabeled dNTPs
are added to the nick - translation mixture , e . g . , DTTP , DATP , and dGTP .
Page 10-9
a - 32P and three unlabeled dNTPs or to dilute each [ a - 32P ] dNTP with an
appropriate amount of the unlabeled dNTP . Most commercial suppliers sell [ a -
32P ] dNTPs in a concentrated aqueous solution , which can be added directly to
the ...
a - 32P and three unlabeled dNTPs or to dilute each [ a - 32P ] dNTP with an
appropriate amount of the unlabeled dNTP . Most commercial suppliers sell [ a -
32P ] dNTPs in a concentrated aqueous solution , which can be added directly to
the ...
Page 10-20
The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al .
1975 ) : dGTP MCTP DATP dTTP 45 um 200 um 12 um 140 uM When the
concentration of each of the four dNTPs in the reaction is greater than Km , the
synthesis ...
The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al .
1975 ) : dGTP MCTP DATP dTTP 45 um 200 um 12 um 140 uM When the
concentration of each of the four dNTPs in the reaction is greater than Km , the
synthesis ...
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Contents
CONCENTRATING NUCLEIC ACIDS E | 8-10 |
SYNTHESIS OF THE FIRST STRAND OF DNA 8 | 8-11 |
MEASUREMENT OF RADIOACTIVITY IN NUCLEIC ACIDS E | 8-18 |
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Common terms and phrases
aliquots allow amount amplification antibody appropriate approximately bacteriophage bacteriophage T4 base binding buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies complete concentration constructed containing deletions denatured described detection determine digestion dNTPs double-stranded efficiency electrophoresis et al ethanol expression extraction Figure filters four fragment gene genomic DNA hybridization increase Incubate inserted interest isolated Klenow labeled length libraries ligation linear linkers membrane method minutes mixture molecules mRNA mutants nucleic acids nucleotides obtained oligonucleotide plaques plasmid DNA plate position possible precipitation prepared presence primer probe problem protein purified radioactivity radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening separate sequence single single-stranded solution step stored strand synthesis target DNA target sequence template termini transfer Tris tube units usually vector volume