Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 15-3
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Generation of Deletions and Insertions SIMPLE DELETIONS OR INSERTIONS The simplest mutations to make are deletions of segments of foreign DNA sequences cloned in plasmid vectors . If the ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Generation of Deletions and Insertions SIMPLE DELETIONS OR INSERTIONS The simplest mutations to make are deletions of segments of foreign DNA sequences cloned in plasmid vectors . If the ...
Page 15-18
... deletions , but also groups of deletions whose endpoints are tightly clustered . • The entire series of enzymatic reactions ( exonuclease digestion , treatment with nuclease S1 or mung - bean nuclease and DNA polymerase , and ...
... deletions , but also groups of deletions whose endpoints are tightly clustered . • The entire series of enzymatic reactions ( exonuclease digestion , treatment with nuclease S1 or mung - bean nuclease and DNA polymerase , and ...
Page 15-33
... deletions must be constructed , sequenced , and matched . For example , in the original description of the method ( McKnight and Kingsbury 1982 ) , 43 5 ' deletions and 42 3 ' deletions were constructed to obtain 15 perfectly matched ...
... deletions must be constructed , sequenced , and matched . For example , in the original description of the method ( McKnight and Kingsbury 1982 ) , 43 5 ' deletions and 42 3 ' deletions were constructed to obtain 15 perfectly matched ...
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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