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TEST LIGATION 3 29
SYNTHESIS OF THE FIRST STRAND OF cDNA 8
COMMONLY USED BUFFERS B
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agarose gel aliquots amino acid amplification antibody appropriate bacterial bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter chromatography Cl pH cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing deletions denatured described digestion dithiothreitol 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 hybridization Incubate the reaction inserted Klenow fragment ligation linear linkers mammalian method MgCl2 microfuge tube minutes at 4°C molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nitrocellulose filter nucleic acids nucleotides oligonucleotide phagemid pipette plaques plasmid plasmid DNA plates polyacrylamide prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature sample screening single-stranded DNA solution specific activity step Store strand of cDNA synthesis T4 DNA ligase target DNA target sequence termini transcription transfer volume