Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-21
... target DNAs into segments that are optimal in size for random sequencing ... DNA in a random fashion that is relatively independent of sequence , and ... DNA is digested to completion with a single restriction enzyme . In theory , sets of ...
... target DNAs into segments that are optimal in size for random sequencing ... DNA in a random fashion that is relatively independent of sequence , and ... DNA is digested to completion with a single restriction enzyme . In theory , sets of ...
Page 13-24
... Target DNA 1. Using the appropriate restriction enzymes , digest a sufficient amount of recombinant vector to yield 10-15 μg of purified target DNA . Separate the target DNA from the vector by preparative gel electrophoresis , and recover ...
... Target DNA 1. Using the appropriate restriction enzymes , digest a sufficient amount of recombinant vector to yield 10-15 μg of purified target DNA . Separate the target DNA from the vector by preparative gel electrophoresis , and recover ...
Page 15-48
A Laboratory Manual Joseph Sambrook, Tom Maniatis. EXCISION OF THE TARGET DNA 1. Excise the target DNA by digesting approximately 50 μg of the recircular- ized plasmid DNA from step 6 , page 15.47 , with the appropriate restric- tion ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. EXCISION OF THE TARGET DNA 1. Excise the target DNA by digesting approximately 50 μg of the recircular- ized plasmid DNA from step 6 , page 15.47 , with the appropriate restric- tion ...
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