Resistance of the cell wall to degradation is a critical parameter for isolation of high quality RNA from natural isolates of Bacillus subtilis
- 111 Downloads
Natural isolates of Bacillus subtilis are known for their ability to produce a large panel of bioactive compounds. Unfortunately, their recalcitrance to conventional molecular techniques limits their transcript studies. In this work, difficulties to isolate RNA attributed to the cell wall were overcome, finally authorising powerful RT-PCR’s.
KeywordsRNA isolation Bacillus subtilis Natural strain Cell wall Transcript analysis RT-PCR
This work received the financial support from the Université des Sciences et Technologies de Lille (BQR), the Agence Nationale de la Recherche (ANR) and the European Funds for the Regional Development. Authors wanted to thank Dr W. Everett for kind re-reading.
- Duitman EH, Hamoen LW, Rembold M, Venema G, Seitz H, Saenger W, Bernhard F, Reinhardt R, Schmidt M, Ullrich C, Stein T, Leenders F, Vater J (1999) The mycosubtilin synthetase of Bacillus subtilis ATCC 6633: a multifunctional hybrid between a peptide synthetase, an amino transferase, and a fatty acid synthase. Proc Natl Acad Sci 96(23):13294–13299PubMedCrossRefGoogle Scholar
- Errington J (1990) Gene cloning techniques. In: Harwood CR, Cutting SM (eds) Molecular biological methods for Bacillus. Wiley, Chichester, pp 175–220Google Scholar
- Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T (2006) The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol 7(3)Google Scholar