Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-31
... primer . In a similar protocol , Han et al . ( 1987 ) use a synthetic primer - adapter consisting of a SalI site and a homopolymeric tract ( 5 ′ GAATTCGTCGACT15 ) to prime the synthesis of the first strand of cDNA . After fractionation ...
... primer . In a similar protocol , Han et al . ( 1987 ) use a synthetic primer - adapter consisting of a SalI site and a homopolymeric tract ( 5 ′ GAATTCGTCGACT15 ) to prime the synthesis of the first strand of cDNA . After fractionation ...
Page 10-20
... primer , template , and dNTPs . The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al . 1975 ) : dGTP 45 μM dCTP 200 μ dATP dTTP 12 μΜ 140 μμ When the concentration of each of the four dNTPs in the reaction is ...
... primer , template , and dNTPs . The Km of the enzyme is different for each of the four dNTPs ( Travaglini et al . 1975 ) : dGTP 45 μM dCTP 200 μ dATP dTTP 12 μΜ 140 μμ When the concentration of each of the four dNTPs in the reaction is ...
Page 13-76
... Primer - binding site deleted from template High concentrations of EDTA present Incorrect primer used Problems with a specific chain - extension / chain - termination mixture Variation in template concentration Dirty templates Dirty ...
... Primer - binding site deleted from template High concentrations of EDTA present Incorrect primer used Problems with a specific chain - extension / chain - termination mixture Variation in template concentration Dirty templates Dirty ...
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Common terms and phrases
agarose gel aliquots amino acid amplification bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA Biol buffer carried cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing deletions denatured digestion DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol ethidium bromide exonuclease formamide gel electrophoresis gene genomic DNA H₂O hybridization Incubate the reaction inserted Klenow fragment labeled ligation linkers method microfuge tube minutes at 4°C mismatched molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nucleotides oligonu oligonucleotide phagemid plaques plasmid plasmid DNA plates polymerase chain reaction pool primer probe protein purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples screening sequencing reactions single-stranded DNA solution specific activity Store strand of cDNA synthesis target DNA target fragment target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml