Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-23
... efficiency of cloning , the number of homopolymeric residues on the plasmid and the cDNA should be approximately equal , with approxi- mately 100 dA / dT residues or 20 dC / dG residues being added to each end of the DNAs ( Peacock et ...
... efficiency of cloning , the number of homopolymeric residues on the plasmid and the cDNA should be approximately equal , with approxi- mately 100 dA / dT residues or 20 dC / dG residues being added to each end of the DNAs ( Peacock et ...
Page 14-30
... efficiency of the amplification process is greatly affected by small differences in any one of a number of variables . These include ( 1 ) the quality and concentrations of the reagents used ( Taq DNA polymerase , dNTPs ...
... efficiency of the amplification process is greatly affected by small differences in any one of a number of variables . These include ( 1 ) the quality and concentrations of the reagents used ( Taq DNA polymerase , dNTPs ...
Page 15-61
... efficiency with which it will be generated . For example , large deletions ( i.e. , deletions of several hundred nucleotides ) are generated with approximately 50 - fold lower efficiency than mutations involving only local changes in ...
... efficiency with which it will be generated . For example , large deletions ( i.e. , deletions of several hundred nucleotides ) are generated with approximately 50 - fold lower efficiency than mutations involving only local changes in ...
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