Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-27
... linkers , and the hairpin loop is cleaved with nuclease S1 and repaired with the Klenow fragment of E. coli DNA polymerase I before linkers of a second type are ligated to the cDNA . These linkers will be added to both ends of the cDNA ...
... linkers , and the hairpin loop is cleaved with nuclease S1 and repaired with the Klenow fragment of E. coli DNA polymerase I before linkers of a second type are ligated to the cDNA . These linkers will be added to both ends of the cDNA ...
Page 15-10
... linker ( in nucleotides ) , y = length of linear plasmid DNA ( in nucleotides ) , and z = desired molar ratio of linkers to target DNA . Usually , a 20- to 30 - fold molar excess of linkers is used to ensure that several copies of the ...
... linker ( in nucleotides ) , y = length of linear plasmid DNA ( in nucleotides ) , and z = desired molar ratio of linkers to target DNA . Usually , a 20- to 30 - fold molar excess of linkers is used to ensure that several copies of the ...
Page 15-85
... linker to the cohesive termini • Removal of excess linkers by cleavage with the appropriate restriction enzyme Ligation of a fragment of DNA carrying a selectable marker ( usually kan ' ) to the cohesive termini of the linkers ...
... linker to the cohesive termini • Removal of excess linkers by cleavage with the appropriate restriction enzyme Ligation of a fragment of DNA carrying a selectable marker ( usually kan ' ) to the cohesive termini of the linkers ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
Copyright | |
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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