Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 48
When using probes that have the capacity to self - anneal ( e . g . , nick -
translated double - stranded DNA ) , the following rule of thumb is useful : Allow
the hybridization to proceed for a time sufficient to enable the probe in solution to
achieve ...
When using probes that have the capacity to self - anneal ( e . g . , nick -
translated double - stranded DNA ) , the following rule of thumb is useful : Allow
the hybridization to proceed for a time sufficient to enable the probe in solution to
achieve ...
Page 53
When 32P - labeled cDNA or RNA is used as a probe , poly ( A ) * RNA at a
concentration of 1 mg / ml may be included in the prehybridization and
hybridization solutions to prevent the probe from binding to T - rich sequences
that are found ...
When 32P - labeled cDNA or RNA is used as a probe , poly ( A ) * RNA at a
concentration of 1 mg / ml may be included in the prehybridization and
hybridization solutions to prevent the probe from binding to T - rich sequences
that are found ...
Page 18
Preparation of Single - stranded Probes Single - stranded probes , which consist
of only one of the two complementary strands of a ... strand eliminates the
possibility of forming nonproductive hybrids composed solely of reannealed
probe .
Preparation of Single - stranded Probes Single - stranded probes , which consist
of only one of the two complementary strands of a ... strand eliminates the
possibility of forming nonproductive hybrids composed solely of reannealed
probe .
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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