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 ΜΜ dATP 12 μ dTTP 140 μ 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 ΜΜ dATP 12 μ dTTP 140 μ 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
acrylamide agarose gel aliquots amplification antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter chromatography cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing ddNTPs deletions denatured described digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers method MgCl2 microfuge tube minutes at 4°C molecules mRNA mutants Natl nitrocellulose nitrocellulose filters nucleic acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing reactions solution specific activity step stored strand of cDNA synthesis target DNA target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml