A monophasic solution for isolation of RNA devoid of polymerase chain reaction-detectable genomic DNA contamination.
Commercially available reagents and published protocols are widely used for RNA isolation. However, genomic DNA contamination in isolated RNA is a potential problem. Here we describe a simple, inexpensive method for eliminating genomic DNA contamination beyond the level of PCR-based detection through reduction of the guanidine thiocyanate concentration (1.5M) in a single monophasic solution based on Chomczynski-Sacchi reagents. The new method can be used to isolate small and large RNA species of high quality and can be completed within an hour.
Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India.
Ujjal
U
Ghosh
Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India.
Shankhamala
S
Bose
Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India.
Trinath
T
Ghosh
Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India.
Tapas Kumar
TK
Bandyopadhyay
Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India. Electronic address: utpal@klyuniv.ac.in.
Utpal
U
Basu
D000818:Animals; D002460:Cell Line; D059018:DNA Contamination; D023281:Genomics; D051379:Mice; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification*
D012313:RNA
Monophasic solution; PCR; RNA