Isolation of mRNA and genomic DNA from epithelial cells in human milk and amplification by PCR

S. Lindquist, L. Hansson, O. Hernell, B. Lonnerdal, J. Normark, M. Stromqvist, S. Bergstrom

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Studies on the regulation of human milk protein genes and suitable cell culture systems have been limited due to restricted availability of tissue samples from lactating women. Although mammary gland tissue has been available from mammary reduction surgery, studies on tissue-specific expression of milk protein genes require samples obtained during specific stages of expression in the tissue, i.e., lactation. We have therefore developed a technique to isolate mRNA and genomic DNA directly from epithelial cells isolated from human milk and used PCR methodology to specifically amplify the cDNA and genomic DNA for human β-casein. When comparing fresh human milk with milk that has been stored for five hours at 37°C or frozen and thawed, we found that the amount of mRNA isolated was considerably higher from fresh milk. However, in spite of this, we could still isolate intact mRNA from frozen milk. The described methodology should be useful in studies on milk protein gene expression during lactation and studies on variants of milk protein genes.

Original languageEnglish (US)
Pages (from-to)692-696
Number of pages5
JournalBioTechniques
Volume17
Issue number4
StatePublished - 1994
Externally publishedYes

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

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    Lindquist, S., Hansson, L., Hernell, O., Lonnerdal, B., Normark, J., Stromqvist, M., & Bergstrom, S. (1994). Isolation of mRNA and genomic DNA from epithelial cells in human milk and amplification by PCR. BioTechniques, 17(4), 692-696.