Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 15-55
... mutagenic oligonucleotide is now free to anneal to incorrect sites on the template . To suppress these effects , between 7 and 9 perfectly matched nucleotides are required at the 3 ' terminus of the mutagenic oligonu- cleotide . • The ...
... mutagenic oligonucleotide is now free to anneal to incorrect sites on the template . To suppress these effects , between 7 and 9 perfectly matched nucleotides are required at the 3 ' terminus of the mutagenic oligonu- cleotide . • The ...
Page 15-57
... mutagenic oligonucleotides of a wide variety of lengths and base compositions . However , annealing mixtures containing mutagenic oligonucleotides that are exceptionally rich in A + T may need to be cooled to lower temperatures ( 12–16 ...
... mutagenic oligonucleotides of a wide variety of lengths and base compositions . However , annealing mixtures containing mutagenic oligonucleotides that are exceptionally rich in A + T may need to be cooled to lower temperatures ( 12–16 ...
Page 15-63
... mutagenic oligonucleotide should be designed as described on pages 15.54-15.56 and should be complementary to the strand of the target DNA that is packaged in bacteriophage M13 particles [ the ( + ) strand ] . Before use in site ...
... mutagenic oligonucleotide should be designed as described on pages 15.54-15.56 and should be complementary to the strand of the target DNA that is packaged in bacteriophage M13 particles [ the ( + ) strand ] . Before use in site ...
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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