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 ... CDNA CLONES OF INTEREST 8 Methods of Screening 8 NUCLEIC ACID HYBRIDIZATION 8.
A Laboratory Manual Joseph Sambrook, Tom Maniatis. IDENTIFICATION OF CDNA CLONES OF INTEREST Methods of Screening There are three methods to screen cDNA ... CDNA CLONES OF INTEREST 8 Methods of Screening 8 NUCLEIC ACID HYBRIDIZATION 8.
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
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acrylamide agarose gel aliquots amplification annealing antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g cleavage cleaved cleotide coli DNA polymerase concentration containing ddNTPs deletions denatured digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers Maxam-Gilbert method MgCl2 microfuge tube minutes at 4°C molecules mRNA Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid piperidine plaques plasmid plasmid DNA plate polymerase chain reaction precipitation prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing gel sequencing reactions sonicated specific activity stored strand of cDNA synthesis T4 DNA polymerase target DNA target sequence template DNA termini transcription transfer Tris Cl pH vectors µg/ml