Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-25
... volume of approximately 0.8 ml . . 6. Neutralize the solution by adding 0.5 volume of 2 M Tris Cl ( pH 8.0 ) and 1 volume of 0.1 N HCl . 7. Check the size of the probe by electrophoresis through a 5 % poly- acrylamide gel ( see Chapter ...
... volume of approximately 0.8 ml . . 6. Neutralize the solution by adding 0.5 volume of 2 M Tris Cl ( pH 8.0 ) and 1 volume of 0.1 N HCl . 7. Check the size of the probe by electrophoresis through a 5 % poly- acrylamide gel ( see Chapter ...
Page 11-38
... volume of 100 μl or less ) on the column . Immediately after the sample has entered the column , add 100 μl of ... volume of phenol : chloroform . Reextract the organic phase with 50 μl of 10 mM Tris Cl ( pH 8.0 ) , and combine the two ...
... volume of 100 μl or less ) on the column . Immediately after the sample has entered the column , add 100 μl of ... volume of phenol : chloroform . Reextract the organic phase with 50 μl of 10 mM Tris Cl ( pH 8.0 ) , and combine the two ...
Page 11-42
... volume as possible . It is therefore better to use as substrates radiolabeled dNTPs supplied in ethanol / water rather than those supplied in buffered aqueous solvents . Appropriate volumes of the ethanolic [ a - 32P ] dNTPs can be ...
... volume as possible . It is therefore better to use as substrates radiolabeled dNTPs supplied in ethanol / water rather than those supplied in buffered aqueous solvents . Appropriate volumes of the ethanolic [ a - 32P ] dNTPs can be ...
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