Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 67
Page 8-6
... interest , or of mRNA that has been partially fragmented by limited alkaline hydrolysis and end - labeled by phosphor- ylation . As a good approximation , the mRNA sequences of interest will be represented in both the probe and the ...
... interest , or of mRNA that has been partially fragmented by limited alkaline hydrolysis and end - labeled by phosphor- ylation . As a good approximation , the mRNA sequences of interest will be represented in both the probe and the ...
Page 8-7
... interest are present at even lower levels ( 1 molecule / cell is not unusual [ Toole et al . 1984 ; Wood et al . 1984 ] ) . Further- more , it is often necessary to clone cDNAs from populations of mRNAs isolated from tissues that ...
... interest are present at even lower levels ( 1 molecule / cell is not unusual [ Toole et al . 1984 ; Wood et al . 1984 ] ) . Further- more , it is often necessary to clone cDNAs from populations of mRNAs isolated from tissues that ...
Page 10-41
... interest cells not expressing mRNA ( s ) of interest poly ( A ) + RNA ann am m Generate radioactive probe by reverse transcription of the entire mRNA population using oligo ( dT ) or short digonucleotides of random sequence as primers ...
... interest cells not expressing mRNA ( s ) of interest poly ( A ) + RNA ann am m Generate radioactive probe by reverse transcription of the entire mRNA population using oligo ( dT ) or short digonucleotides of random sequence as primers ...
Contents
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 8-11 |
MOLECULAR CLONING OF DOUBLESTRANDED cDNA 8 | 8-21 |
8 | 8-29 |
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Common terms and phrases
acrylamide agarose gel aliquots amplification antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter chromatography cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing ddNTPs deletions denatured described digestion diluted dithiothreitol DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers method MgCl2 microfuge tube minutes at 4°C molecules mRNA mutants Natl nitrocellulose nitrocellulose filters nucleic acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing reactions solution specific activity step stored strand of cDNA synthesis target DNA target sequence template DNA termini transcription transfer Tris Cl pH vector