Lung tissue storage: optimizing conditions for future use in molecular research.
The quality of tissue studied impacts greatly on oligonucleotide microarray results, emphasizing the importance of harvesting techniques. The analyzed RNA extracted from human lung samples preserved via 4 different storage conditions (RNAlater, phosphate-buffered saline, TRIzol, liquid nitrogen). RNA was assessed by denaturing gel electrophoresis, Agilent bioanalysis, real-time polymerase chain reaction (PCR), and Test3 Affymetrix chip hybridization. Results revealed better quality RNA from RNAlater samples on gel electrophoresis and bioanalysis. RNAlater samples also showed greater yield (r18s via PCR P < .05) and resulted in better Test3 chips hybridization (p < .05), suggesting RNAlater was superior at preserving lung tissue nucleic acid.
General Clinical Research Unit, School of Medicine and Medical Sciences, University College Dublin, Mater Misericordiae University Hospital, and Dublin Molecular Medicine Centre, Dublin, Ireland.
Dominic T K
DT
Keating
Andrea P
AP
Malizia
D
D
Sadlier
C
C
Hurson
A E
AE
Wood
J
J
McCarthy
L
L
Nolke
Jim J
JJ
Egan
Peter P
PP
Doran
D000368:Aged; D004586:Electrophoresis; D005260:Female; D006801:Humans; D008168:Lung; D008297:Male; D046228:Microarray Analysis / Q000379:methods; D008875:Middle Aged; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000627:therapeutic use; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis
D019188:Organ Preservation Solutions; D012313:RNA