Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-16
However , if it is necessary to clone the 5 ' - terminal sequences of eukaryotic
mRNA with high efficiency , several more - difficult methods are available . For
example , after completion of first - strand synthesis , terminal transferase
transferase ...
However , if it is necessary to clone the 5 ' - terminal sequences of eukaryotic
mRNA with high efficiency , several more - difficult methods are available . For
example , after completion of first - strand synthesis , terminal transferase
transferase ...
Page 8-23
For optimal efficiency of cloning , the number of homopolymeric residues on the
plasmid and the cDNA should be approximately equal , with approximately 100
dA / dT residues or 20 dC / G residues being added to each end of the DNAs ...
For optimal efficiency of cloning , the number of homopolymeric residues on the
plasmid and the cDNA should be approximately equal , with approximately 100
dA / dT residues or 20 dC / G residues being added to each end of the DNAs ...
Page 15-61
Methods to Improve the Efficiency of Oligonucleotide - mediated Mutagenesis
The basic procedures described above have been used ... The larger and more
complex the mutation , the lower the efficiency with which it will be generated .
Methods to Improve the Efficiency of Oligonucleotide - mediated Mutagenesis
The basic procedures described above have been used ... The larger and more
complex the mutation , the lower the efficiency with which it will be generated .
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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