Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-34
... template DNA by digestion with a suitable restriction enzyme . Analyze an aliquot ( 100 ng ) of the digested DNA by agarose gel electrophoresis . If necessary , add more restriction enzyme and continue incubation until there is no trace ...
... template DNA by digestion with a suitable restriction enzyme . Analyze an aliquot ( 100 ng ) of the digested DNA by agarose gel electrophoresis . If necessary , add more restriction enzyme and continue incubation until there is no trace ...
Page 13-7
... DNA can be used as templates in the Sanger method of sequencing : pure single - stranded DNA and double - strand- ed ... template to primer , it is possible to obtain several hundred nucleotides of sequence from each set of chain- termination ...
... DNA can be used as templates in the Sanger method of sequencing : pure single - stranded DNA and double - strand- ed ... template to primer , it is possible to obtain several hundred nucleotides of sequence from each set of chain- termination ...
Page 15-74
... TEMPLATE STRANDS THAT CONTAIN URACIL ( KUNKEL METHOD ) This method ( Kunkel 1985 ; Kunkel et al . 1987 ) uses single ... DNA at sites normally occupied by thymine . E. coli normally synthesizes an enzyme ( uracil - N - glycosylase ) that ...
... TEMPLATE STRANDS THAT CONTAIN URACIL ( KUNKEL METHOD ) This method ( Kunkel 1985 ; Kunkel et al . 1987 ) uses single ... DNA at sites normally occupied by thymine . E. coli normally synthesizes an enzyme ( uracil - N - glycosylase ) that ...
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