Molecular Cloning: A Laboratory Manual, Book 2 |
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Page xvi
A Laboratory Manual Joseph Sambrook, Tom Maniatis. 8 Construction and Analysis of cDNA Libraries Strategies for cDNA Cloning 8.3 PREPARATION OF mRNA FOR CDNA CLONING 8.3 Source of the mRNA 8.3 Integrity of the mRNA 8.4 Abundant mRNAs 8.6 ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. 8 Construction and Analysis of cDNA Libraries Strategies for cDNA Cloning 8.3 PREPARATION OF mRNA FOR CDNA CLONING 8.3 Source of the mRNA 8.3 Integrity of the mRNA 8.4 Abundant mRNAs 8.6 ...
Page 8-7
... clones required to achieve a given probability that a low - abundance mRNA will be present in a cDNA library is N = In ( 1 - P ) In ( 1-1 / n ) where N = the number of clones required , P = the probability desired ( usually 0.99 ) , and ...
... clones required to achieve a given probability that a low - abundance mRNA will be present in a cDNA library is N = In ( 1 - P ) In ( 1-1 / n ) where N = the number of clones required , P = the probability desired ( usually 0.99 ) , and ...
Page 8-46
A Laboratory Manual Joseph Sambrook, Tom Maniatis. IDENTIFICATION OF CDNA CLONES OF INTEREST Methods of Screening There are three methods to screen cDNA libraries for clones of interest : • Nucleic acid hybridization ... cDNA CLONES 8 8.
A Laboratory Manual Joseph Sambrook, Tom Maniatis. IDENTIFICATION OF CDNA CLONES OF INTEREST Methods of Screening There are three methods to screen cDNA libraries for clones of interest : • Nucleic acid hybridization ... cDNA CLONES 8 8.
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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 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 µg/ml