Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-58
... Cl ( pH 7.4 ] ) . Store in aliquots at -70 ° C . Packaging extracts are much cheaper to make than to buy . If you plan to construct more than one or two cDNA libraries , it would be worthwhile making a large batch of packaging extract ...
... Cl ( pH 7.4 ] ) . Store in aliquots at -70 ° C . Packaging extracts are much cheaper to make than to buy . If you plan to construct more than one or two cDNA libraries , it would be worthwhile making a large batch of packaging extract ...
Page 8-59
... pH 5.2 ) Synthetic linkers TE ( pH 7.6 and pH 8.0 ) Trichloroacetic acid ( TCA ) ( 10 % ) Tris base ( 2 M ) Tris Cl ( 2 м ; pH 7.4 ) · Tris Cl ( 1 м ; pH 7.6 ) Tris Cl ( 2 м ; pH 8.0 ) Tris Cl ( 1 м ; pH 8.3 ) X - gal Add Sephadex G ...
... pH 5.2 ) Synthetic linkers TE ( pH 7.6 and pH 8.0 ) Trichloroacetic acid ( TCA ) ( 10 % ) Tris base ( 2 M ) Tris Cl ( 2 м ; pH 7.4 ) · Tris Cl ( 1 м ; pH 7.6 ) Tris Cl ( 2 м ; pH 8.0 ) Tris Cl ( 1 м ; pH 8.3 ) X - gal Add Sephadex G ...
Page 8-68
... Cl ( pH 8.3 ) 33 mM MgCl2 50 mm ẞ - mercaptoethanol This buffer should be stored in small aliquots at −20 ° C . Add 1-2 units of bacteriophage T4 DNA polymerase ( 500 units / ml ) and incubate for 15 minutes at 37 ° C . 3. Stop the ...
... Cl ( pH 8.3 ) 33 mM MgCl2 50 mm ẞ - mercaptoethanol This buffer should be stored in small aliquots at −20 ° C . Add 1-2 units of bacteriophage T4 DNA polymerase ( 500 units / ml ) and incubate for 15 minutes at 37 ° C . 3. Stop the ...
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