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  • Title: Single-cell cDNA-PCR: removal of contaminating genomic DNA from total RNA using immobilized DNase I.
    Author: Ziegler BL, Lamping C, Thoma S, Thomas CA.
    Journal: Biotechniques; 1992 Nov; 13(5):726-9. PubMed ID: 1418973.
    Abstract:
    A procedure utilizing immobilized DNase I that allows the efficient amplification of cDNA by PCR from a single cell in the absence of contaminating genomic DNA is described. DNase I treated, total RNA derived from single cells was reverse transcribed into cDNA followed by PCR using beta-actin and c-fos specific primers that recognize different exons of the respective genes. Amplification products corresponding to cDNA, but not to genomic sequences, were detected after treatment with immobilized DNase I in samples previously shown to be contaminated with genomic DNA. This method allows the efficient removal of DNA contaminating total RNA derived from a single cell.
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