Abstract
A rapid and reliable procedure for molecular cloning and nucleotide sequencing of PCR product is described. PCR amplified fragment recovered from agarose gel by centrifugation through siliconized sterilized glass wool was further puri fled in sequential extraction with phenol, phenol/chloroform and chloroform. This was ligated into Thanged plasmid vector and transformed E. Coli was isolated. The plasmid recovered by the alkali denature method was purified by means of spun columns containing Sepharose CL-4B. This step is critical in obtaining a reading >500 by with an automatic DNA sequencer.
