Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 6
In these cases , the RNA species of interest constitutes as much as 50 - 90 % of
the total poly ( A ) * cytoplasmic RNA isolated from specific types of differentiated
cells . Consequently , no further purification of the particular mRNA is required ...
In these cases , the RNA species of interest constitutes as much as 50 - 90 % of
the total poly ( A ) * cytoplasmic RNA isolated from specific types of differentiated
cells . Consequently , no further purification of the particular mRNA is required ...
Page 7
99 1 / n = 1 / 37 , 000 N = 170 , 000 Unfortunately , many mRNAs of interest are
present at even lower levels ( 1 molecule / cell is not unusual [ Toole et al . 1984 ;
Wood et al . 1984 ] ) . Furthermore , it is often necessary to clone cDNAs from ...
99 1 / n = 1 / 37 , 000 N = 170 , 000 Unfortunately , many mRNAs of interest are
present at even lower levels ( 1 molecule / cell is not unusual [ Toole et al . 1984 ;
Wood et al . 1984 ] ) . Furthermore , it is often necessary to clone cDNAs from ...
Page 41
avovovovova cells expressing mRNA ( s ) of interest cells not expressing mRNA (
s ) of interest poly ( A ) * RNA poly ( A ) * RNA Generate radioactive probe by
reverse transcription of the entire mRNA population using oligo ( dT ) or short ...
avovovovova cells expressing mRNA ( s ) of interest cells not expressing mRNA (
s ) of interest poly ( A ) * RNA poly ( A ) * RNA Generate radioactive probe by
reverse transcription of the entire mRNA population using oligo ( dT ) or short ...
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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 |
Copyright | |
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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