Cloning of PCR synthesized 191 bp DNA overlapping lac promoter for plasmid construction.
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1997-01-01
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E coli genomic DNA was amplified by polymerase chain reaction (PCR) using two 5' and 3' oligonucleotide primers (27-mer). Amplified DNA was 191 bp. The region of amplified DNA on lac operon in E coli was 14 bp upstream from the transcription initiation site and 177 bp downstream. Amplified DNA was cloned into a phagemid vector for construction of plasmid, suitable for use as template for making strand-specific probe to detect initiated lac transcript by RNase protection assay, labelling for Southern and Northern hybridization. Another use of this probe made from this construct is to detect strand-specific DNA repair. The construct was verified by DNA sequencing.
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Biswas SR. Cloning of PCR synthesized 191 bp DNA overlapping lac promoter for plasmid construction. Indian Journal of Experimental Biology. 1997 Jan; 35(1): 99-102