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Complementary  DNA (cDNA) is valuable for investigating protein structure and function in the  study of life science, but it is difficult to obtain by traditional reverse  transcription. We employed a novel strategy to clone human leukemia inhibitory  factor (hLIF) gene cDNA from genomic DNA, which was directly isolated from the  mucous membrane of mouth. The hLIF sequence, which is 609 bp long and is  composed of three exons, can be acquired within a few hours by amplifying each  exon and splicing all of them using overlap-PCR. This new approach developed is  simple, time- and cost-effective, without RNA preparation or cDNA synthesis,  and is not limited to the specific tissues for a particular gene and the  expression level of the gene.

A new approach for cloning hLIF cDNA from genomic DNA isolated from the oral mucous membrane