Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 62
... 33 ug = ug of first strand of cDNA synthesized total cpm iii . A reaction mixture
buffered at pH 8 . 3 is generally used to synthesize cDNA with reverse
transcriptase purified from particles of avian retroviruses : poly ( A ) * RNA ( 1 mg /
ml ) 10 .
... 33 ug = ug of first strand of cDNA synthesized total cpm iii . A reaction mixture
buffered at pH 8 . 3 is generally used to synthesize cDNA with reverse
transcriptase purified from particles of avian retroviruses : poly ( A ) * RNA ( 1 mg /
ml ) 10 .
Page 13
Purification of Antisera When the antisera to be used for screening react with only
one protein of the expected molecular ... However , if the antibodies that react
with the antigen of interest are of the IgG class , they may be easily purified and ...
Purification of Antisera When the antisera to be used for screening react with only
one protein of the expected molecular ... However , if the antibodies that react
with the antigen of interest are of the IgG class , they may be easily purified and ...
Page 24
GENERATION OF A LIBRARY OF RANDOMLY OVERLAPPING CLONES
Purification and Ligation of the Target DNA 1 . Using the appropriate restriction
enzymes , digest a sufficient amount of recombinant vector to yield 10 – 15 ug of
purified ...
GENERATION OF A LIBRARY OF RANDOMLY OVERLAPPING CLONES
Purification and Ligation of the Target DNA 1 . Using the appropriate restriction
enzymes , digest a sufficient amount of recombinant vector to yield 10 – 15 ug of
purified ...
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Contents
TEST LIGATION 3 29 | 3 |
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 11 |
FRAGMENT OF E coli DNA POLYMERASE I 13 59 | 14 |
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Common terms and phrases
acid activity agarose aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies concentration constructed containing deletions denatured described determine digestion DNA polymerase dNTPs double-stranded efficiency electrophoresis et al ethanol expression Figure filters fragment gene genomic hybridization increase Incubate inserted interest isolated labeled length libraries ligation linear linkers method microfuge minutes mixture molecules mRNA mutagenesis mutagenic mutations Nucleic nucleotides obtained oligonucleotide original plaques plasmid plasmid DNA plate pool position possible prepared presence primer probe problem protein purified radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening sequence single single-stranded solution specific step Store strand synthesis target DNA target sequence template termini tion transfer transformants tube units usually vector volume