Optimization of RNA preservation in granulation tissue in the expanded polytetrafluoroethylene (ePTFE) tube model.
RNA is easily degraded by RNAses. An efficient and non-hazardous method optimizing RNA preservation and extraction is needed. Our aim was to evaluate RNA preservation methods on murine granulation tissue from subcutaneously implanted ePTFE tubes. ePTFE tubes were placed subcutaneously in mice, removed after ten days and randomized to four RNA preservation groups. Storage in RNAlater at 4 degrees C and snap freezing in methanol/acetone yielded significantly better RNA quality than snap freezing with liquid nitrogen and storage in RNAlater at -80 degrees C. Snap freezing in liquid nitrogen failed to yield intact RNA. We recommend storage in RNAlater at 4 degrees C.
Danielsen
Patricia L
PL
Department of Surgery K, Bispebjerg Hospital, University of Copenhagen, Bispebjerg Bakke 23, Copenhagen, Denmark. patriciadanielsen@yahoo.dk
Koudahl
V
V
Christensen
B
B
Olsen
J
J
D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D005465:Fluorocarbon Polymers; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D013048:Specimen Handling / Q000379:methods*; D014945:Wound Healing / Q000235:genetics*
D005465:Fluorocarbon Polymers; C010946:polytetrafluoroethane; D012313:RNA