Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 24
Because the next step of the procedure requires the target DNA to be ligated to
itself , it is best to use restriction enzymes that generate compatible termini . If this
is not possible , an acceptable alternative is to use restriction enzymes that ...
Because the next step of the procedure requires the target DNA to be ligated to
itself , it is best to use restriction enzymes that generate compatible termini . If this
is not possible , an acceptable alternative is to use restriction enzymes that ...
Page 32
3000 Ci / mmole ) 10 uCi It is best to generate this series of dilutions from the
appropriate original tenfold dilution of the control template ( step 2 ) . Do not
reduce the concentration of unlabeled dCTP in the reaction mixture to increase
the ...
3000 Ci / mmole ) 10 uCi It is best to generate this series of dilutions from the
appropriate original tenfold dilution of the control template ( step 2 ) . Do not
reduce the concentration of unlabeled dCTP in the reaction mixture to increase
the ...
Page 46
The efficiency of transformation drops markedly when more than 200 ul of
competent cells are used per reaction , probably because the timing of the heat -
shock step is no longer optimal . 12 . Just before plating on selective media , pool
the ...
The efficiency of transformation drops markedly when more than 200 ul of
competent cells are used per reaction , probably because the timing of the heat -
shock step is no longer optimal . 12 . Just before plating on selective media , pool
the ...
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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