Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 23
The reaction often proceeds asynchronously , so that different termini of DNA
molecules carry homopolymeric tails of very different lengths . This problem can
be alleviated , but not completely solved , by using plasmids carrying ...
The reaction often proceeds asynchronously , so that different termini of DNA
molecules carry homopolymeric tails of very different lengths . This problem can
be alleviated , but not completely solved , by using plasmids carrying ...
Page 51
Labeling the 5 ' and 3 ' Termini of DNA The termini of DNA molecules can be
radiolabeled enzymatically to yield molecules that may be used in nuclease - Si
mapping of RNA , as radiolabeled primers in primer - extension reactions , and
as ...
Labeling the 5 ' and 3 ' Termini of DNA The termini of DNA molecules can be
radiolabeled enzymatically to yield molecules that may be used in nuclease - Si
mapping of RNA , as radiolabeled primers in primer - extension reactions , and
as ...
Page 60
Forward Reaction USING DNA MOLECULES WITH PROTRUDING 5 ' TERMINI
AS SUBSTRATES 1 . Mix : 5 ' termini of dephosphorylated DNA ( see pages 10 .
64 – 10 . 65 ) 1 - 50 pmoles 10 X bacteriophage T4 polynucleotide kinase buffer ...
Forward Reaction USING DNA MOLECULES WITH PROTRUDING 5 ' TERMINI
AS SUBSTRATES 1 . Mix : 5 ' termini of dephosphorylated DNA ( see pages 10 .
64 – 10 . 65 ) 1 - 50 pmoles 10 X bacteriophage T4 polynucleotide kinase buffer ...
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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