Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 14
The specific activity of the product depends on the amount of template added to
the reaction : the lower the amount of template , the greater the specific activity .
Under the reaction conditions described , radiolabeling to maximum specific ...
The specific activity of the product depends on the amount of template added to
the reaction : the lower the amount of template , the greater the specific activity .
Under the reaction conditions described , radiolabeling to maximum specific ...
Page 43
This method has been used successfully to construct cDNA libraries that are
specific for the gastrulation stage of Xenopus development ( Sargent and Dawid
1983 ) , for murine T lymphocytes ( Davis et al . 1984 ; Hedrick et al . 1984 ) , and
for ...
This method has been used successfully to construct cDNA libraries that are
specific for the gastrulation stage of Xenopus development ( Sargent and Dawid
1983 ) , for murine T lymphocytes ( Davis et al . 1984 ; Hedrick et al . 1984 ) , and
for ...
Page 41
specific activities as high as 2 x 10 " cpm per microgram of probe . In a
modification of the procedure , longer probes can be synthesized from two
oligonucleotides that contain complementary sequences at their 3 ' termini (
Ullrich et al .
specific activities as high as 2 x 10 " cpm per microgram of probe . In a
modification of the procedure , longer probes can be synthesized from two
oligonucleotides that contain complementary sequences at their 3 ' termini (
Ullrich et al .
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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