Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-88
... cleavage reactions are carried out on one day and the gels are run on the following day . However , with practice , it is possible to complete both cleavage and electrophoresis in a single working day , if a sequencing gel is prepared ...
... cleavage reactions are carried out on one day and the gels are run on the following day . However , with practice , it is possible to complete both cleavage and electrophoresis in a single working day , if a sequencing gel is prepared ...
Page 13-99
... cleavage . Purify restriction enzyme , phosphatase , ki- nase , or terminal transferase . After labeling ragged ends , cleave with re- striction enzymes that will give end seg- ments less than 100 bp long , denature , and fractionate on ...
... cleavage . Purify restriction enzyme , phosphatase , ki- nase , or terminal transferase . After labeling ragged ends , cleave with re- striction enzymes that will give end seg- ments less than 100 bp long , denature , and fractionate on ...
Page 13-100
... cleavage reactions that do not normally share base specificity . Cleavage is consistent with base specificity every- where else in the sequencing pattern . 13. Difficulty in identifying the band corre- sponding to chemical cleavage of ...
... cleavage reactions that do not normally share base specificity . Cleavage is consistent with base specificity every- where else in the sequencing pattern . 13. Difficulty in identifying the band corre- sponding to chemical cleavage of ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
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acrylamide agarose gel aliquots amplification annealing antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g cleavage cleaved cleotide coli DNA polymerase concentration containing ddNTPs deletions denatured digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers Maxam-Gilbert method MgCl2 microfuge tube minutes at 4°C molecules mRNA Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid piperidine plaques plasmid plasmid DNA plate polymerase chain reaction precipitation prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing gel sequencing reactions sonicated specific activity stored strand of cDNA synthesis T4 DNA polymerase target DNA target sequence template DNA termini transcription transfer Tris Cl pH vectors µg/ml