Impact of purified water quality on molecular biology experiments.
Purified water is a reagent used in a variety of molecular biology experiments, for sample and media preparation, in mobile phases of liquid chromatography techniques, and in rinsing steps. The combination of several technologies in water purification systems allows delivering high-purity water adapted to each application and technique. Through a series of examples, the importance of water quality on biotechnology experiments, such as single nucleotide polymorphism (SNP) analysis by denaturating HPLC, RNA preparation and PCR, is presented. Results obtained on DNA mutation and single nucleotide polymorphism analysis using the denaturating HPLC (DHPLC) technique highlight the benefits of organic removal by UV photooxidation process. Comparative gel electrophoresis data show that ultrafiltration is as efficient as diethylpyrocarbonate (DEPC) treatment for suppressing RNase activity in water. Gel electrophoresis and densitometry measurement also point out the benefits of ultrafiltration to carry out reverse transcriptase-polymerase chain reaction quantitatively.
Research and Development, Lab Water Division, Millipore S.A.S., St Quentin-Yvelines, France. stephane_mabic@millipore.com
Stéphane
S
Mabic
Ichiro
I
Kano
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology*; D010179:Pancreas / Q000737:chemistry / Q000201:enzymology; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012313:RNA / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*; D014462:Ultrafiltration; D014867:Water / Q000737:chemistry
D014867:Water; D012313:RNA; D012259:Ribonuclease, Pancreatic; D004047:Diethyl Pyrocarbonate