Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-80
A Laboratory Manual Joseph Sambrook, Tom Maniatis. PROBLEMS COMMONLY ENCOUNTERED WITH cDNA CLONING Problems commonly ... problem still persists , check the other components of the first - strand reaction , using an mRNA template whose ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. PROBLEMS COMMONLY ENCOUNTERED WITH cDNA CLONING Problems commonly ... problem still persists , check the other components of the first - strand reaction , using an mRNA template whose ...
Page 13-74
... problems are listed in Table 13.5 together with suggestions for solving them . When problems of this type arise , it is ... problem persists , try substituting the base analog dITP for dGTP in the chain - extension / chain - termination ...
... problems are listed in Table 13.5 together with suggestions for solving them . When problems of this type arise , it is ... problem persists , try substituting the base analog dITP for dGTP in the chain - extension / chain - termination ...
Page 15-80
... problem regions . Use a shorter oligonucleotide as a hybridization probe to screen for the desired mutant . 3. All plaques or colonies hybridize to the oligonucleotide probe . • Check that there is no region of extensive homology ...
... problem regions . Use a shorter oligonucleotide as a hybridization probe to screen for the desired mutant . 3. All plaques or colonies hybridize to the oligonucleotide probe . • Check that there is no region of extensive homology ...
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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