Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-11
... reaction and that the enzyme may be sensitive to contaminants found in ... Incubate for 15 minutes at 37 ° C . Each new batch of DNAase I should be ... reaction on ice and add : 10 nick - translation buffer ( see page 10.7 ) unlabeled ...
... reaction and that the enzyme may be sensitive to contaminants found in ... Incubate for 15 minutes at 37 ° C . Each new batch of DNAase I should be ... reaction on ice and add : 10 nick - translation buffer ( see page 10.7 ) unlabeled ...
Page 10-22
... reaction by tapping the side of the tube . Incubate the reaction for 30 minutes at room temperature . 6. Transfer 0.5 μl of the reaction to a fresh microfuge tube containing 20 μl of 0.5 M EDTA ( pH 8.0 ) . Store the tube on ice . 7 ...
... reaction by tapping the side of the tube . Incubate the reaction for 30 minutes at room temperature . 6. Transfer 0.5 μl of the reaction to a fresh microfuge tube containing 20 μl of 0.5 M EDTA ( pH 8.0 ) . Store the tube on ice . 7 ...
Page 10-55
... incubate for the appropriate time . If the desired restriction enzyme does not function in bacteriophage T4 DNA ... reaction for 5 minutes at 37 ° C . 7. Add 1 μl of a 2 mm solution of the unlabeled fourth dNTP . Continue the incubation for a ...
... incubate for the appropriate time . If the desired restriction enzyme does not function in bacteriophage T4 DNA ... reaction for 5 minutes at 37 ° C . 7. Add 1 μl of a 2 mm solution of the unlabeled fourth dNTP . Continue the incubation for a ...
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