Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-24
... cleaved vector with a phosphatase before ligation to cDNA . If the double - stranded cDNA contains one or more recognition sites for the restriction enzyme , it will be cleaved and subsequently cloned as two or more DNA fragments ...
... cleaved vector with a phosphatase before ligation to cDNA . If the double - stranded cDNA contains one or more recognition sites for the restriction enzyme , it will be cleaved and subsequently cloned as two or more DNA fragments ...
Page 8-27
... cleaved . The linkers are therefore added only to one end of the double - stranded cDNA ( the end corresponding to the 3 ' terminus of the mRNA ) . The cDNA is then purified from excess unligated linkers , and the hairpin loop is cleaved ...
... cleaved . The linkers are therefore added only to one end of the double - stranded cDNA ( the end corresponding to the 3 ' terminus of the mRNA ) . The cDNA is then purified from excess unligated linkers , and the hairpin loop is cleaved ...
Page 13-99
... cleaved at adenines carry too many ring - opened 7- methylguanines and are heterogeneous in charge . Sample not evenly distributed in sample well . Nested end - labeled cleavage products be- come long enough to fold into stable hair ...
... cleaved at adenines carry too many ring - opened 7- methylguanines and are heterogeneous in charge . Sample not evenly distributed in sample well . Nested end - labeled cleavage products be- come long enough to fold into stable hair ...
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