Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 16
Page 8-39
... ( HindIII / BamHI / EcoRI ) at codons 5-10 of the lacZ gene ( see map of AORF8 in Chapter 2 , page 2.50 ) . Insertion ... HindIII . Because M.HindIII methylase is not commercially available , the cDNA is methylated with M.BamHI methylase ...
... ( HindIII / BamHI / EcoRI ) at codons 5-10 of the lacZ gene ( see map of AORF8 in Chapter 2 , page 2.50 ) . Insertion ... HindIII . Because M.HindIII methylase is not commercially available , the cDNA is methylated with M.BamHI methylase ...
Page 8-40
... HindIII and BamHI 5 ' pGATC 3 ' 3 ' A AGCTp 5 ' HindIII BamHI left arm right arm lacy lacz lacPO lacl amp ' transcription FIGURE 8.13 Directional cloning of cDNA into AORF8 . bacteriophage λgt22 Not ! - Sall- T 5 ' cap. 8.40 Construction ...
... HindIII and BamHI 5 ' pGATC 3 ' 3 ' A AGCTp 5 ' HindIII BamHI left arm right arm lacy lacz lacPO lacl amp ' transcription FIGURE 8.13 Directional cloning of cDNA into AORF8 . bacteriophage λgt22 Not ! - Sall- T 5 ' cap. 8.40 Construction ...
Page 8-41
... HindIII site is formed by fusion of the poly ( dT : dA ) tract at the 3 ' terminus of the cDNA to the first linker ( see Figure 8.13 ) . Addition of linkers to the 5 ' terminus of the cDNA does not usually create a new restriction site ...
... HindIII site is formed by fusion of the poly ( dT : dA ) tract at the 3 ' terminus of the cDNA to the first linker ( see Figure 8.13 ) . Addition of linkers to the 5 ' terminus of the cDNA does not usually create a new restriction site ...
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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