PaxGene Blood RNA Kit Handbook - Plastisch chirurgen

Appendix A: General Remarks on Handling RNAHandling RNARibonucleases (RNases) are very stable and active enzymes that generally do notrequire cofactors to function. Since RNases are difficult to inactivate and even minuteamounts are sufficient to destroy RNA, do not use any plasticware or glasswarewithout first eliminating possible RNase contamination. Great care should be taken toavoid inadvertently introducing RNases into the RNA sample during or after thepurification procedure. In order to create and maintain an RNase-free environment,precautions must be taken during pretreatment and use of disposable and nondisposablevessels and solutions while working with RNA.General handlingProper microbiological, aseptic technique should always be used when working withRNA. Hands and dust particles carry bacteria and molds and are the most commonsources of RNase contamination. Always wear latex or vinyl gloves while handlingreagents and RNA samples to prevent RNase contamination from the surface of theskin or from dusty laboratory equipment. Change gloves frequently and keep tubesclosed whenever possible. Keep purified RNA on ice when aliquots are pipetted fordownstream applications.Protocols for removing RNase-contamination from glassware and solutions can befound in general molecular biology guides, such as Sambrook, J. and Russell, D. W.(2001) Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor, NY:Cold Spring Harbor Laboratory Press.PAXgene Blood RNA Kit Handbook 06/2005 29