Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 15-52
... desired mutation by oligonucleotide hybridization ( Wallace et al . 1979 , 1981b ) , ( 4 ) the development of a versatile set of bacteriophage M13 vectors ( for review , see Chapter 4 ) , and ( 5 ) the invention of genetic methods to ...
... desired mutation by oligonucleotide hybridization ( Wallace et al . 1979 , 1981b ) , ( 4 ) the development of a versatile set of bacteriophage M13 vectors ( for review , see Chapter 4 ) , and ( 5 ) the invention of genetic methods to ...
Page 15-54
... desired site and the length to which the primer has been extended ( Zoller and Smith 1984 ) . However , tests of this type are generally carried out only when difficulties are encountered in obtaining the desired mutation . The design ...
... desired site and the length to which the primer has been extended ( Zoller and Smith 1984 ) . However , tests of this type are generally carried out only when difficulties are encountered in obtaining the desired mutation . The design ...
Page 15-80
... desired mutation . • Purify the mutagenic oligonucleotide by polyacrylamide gel electro- phoresis . • Test the mutagenic oligonucleotide for its ability to prime DNA synthe- sis at the appropriate location on the single - stranded ...
... desired mutation . • Purify the mutagenic oligonucleotide by polyacrylamide gel electro- phoresis . • Test the mutagenic oligonucleotide for its ability to prime DNA synthe- sis at the appropriate location on the single - stranded ...
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Common terms and phrases
agarose gel aliquots amplification annealing antibody bacterial bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase concentration containing deletions denatured digestion dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Incubate inserted Klenow fragment ligation linear linkers method microfuge tube minutes at 4°C molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probes purified radioactivity radiolabeled reaction mixture recombinant remove restriction enzyme reverse transcriptase room temperature samples screening sequencing reactions single-stranded DNA solution step Store strand of cDNA synthesis target DNA target sequence template DNA termini transfer Tris Cl pH vector