Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 55
Most losses of cDNA occur during precipitation with ethanol . Taking the following
precautions helps to avoid this problem : • After adjusting the salt concentration of
the DNA solution to 0 . 3 m sodium acetate or 2 m ammonium acetate and ...
Most losses of cDNA occur during precipitation with ethanol . Taking the following
precautions helps to avoid this problem : • After adjusting the salt concentration of
the DNA solution to 0 . 3 m sodium acetate or 2 m ammonium acetate and ...
Page 22
Using wide - bore pipettes , carefully layer the cell suspensions under the ethanol
. 5 . Recover the DNA from each tube by slowly stirring the interface between the
cell lysate and the ethanol with a pasteur pipette whose end has been sealed ...
Using wide - bore pipettes , carefully layer the cell suspensions under the ethanol
. 5 . Recover the DNA from each tube by slowly stirring the interface between the
cell lysate and the ethanol with a pasteur pipette whose end has been sealed ...
Page 98
Large , water - insoluble pellets after the first ethanol precipitation e . Poor
suppression of Ts in the C ladder If magnesium acetate is still used in the
hydrazine stop solution as originally suggested , delete it . If piperidine reactions
are done in ...
Large , water - insoluble pellets after the first ethanol precipitation e . Poor
suppression of Ts in the C ladder If magnesium acetate is still used in the
hydrazine stop solution as originally suggested , delete it . If piperidine reactions
are done in ...
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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