Preservation of nucleic acids for polymerase chain reaction after prolonged storage at room temperature.
A guanidinium isothiocyanate (GITC) lysis solution was evaluated for its efficacy in preserving nucleic acids for subsequent analysis after prolonged storage at room temperature. Aliquots of thyroid crude cell lysates were stored at 22 degrees C for 8 weeks in the GITC solution. Crude cell lysates stored for 2 weeks in the GITC solution consistently provided adequate RNA and DNA for reverse transcriptase-polymerase chain reaction (RT-PCR) and PCR, respectively, of beta 2-microglobulin, thyroid stimulating hormone receptor (TSHR), and thyroglobulin. Interestingly, the thyroglobulin and TSHR messages from thyroid fine needle aspirate samples were detected by RT-PCR only when stored at room temperature for less than 1 week, whereas the RT-PCR product for beta 2-microglobulin was detectable in 95% of the samples at 3 months. In addition, thyroglobulin DNA was amplified by PCR in nearly all samples stored at 22 degrees C for 2 months. These data suggest that GITC solutions can be used to preserve nucleic acids in most circumstance until transported to laboratories equipped and staffed with appropriate resources.
Tuttle
R M
RM
Department of Clinical Investigation, Walter Reed Army Medical Center, Washington, DC 20307-5001, USA.
Waselenko
J K
JK
Yosseffi
P
P
Weigand
N
N
Martin
R K
RK
D001707:Biopsy, Needle; D002021:Buffers; D002458:Cell Fractionation / Q000379:methods; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D006146:Guanidines; D006801:Humans; D017879:Isothiocyanates; D011309:Preservation, Biological / Q000379:methods*; D012333:RNA, Messenger / Q000737:chemistry* / Q000302:isolation & purification; D011989:Receptors, Thyrotropin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012996:Solutions; D013053:Spectrophotometry; D013696:Temperature; D013954:Thyroglobulin / Q000235:genetics; D013961:Thyroid Gland / Q000737:chemistry / Q000473:pathology; D001613:beta 2-Microglobulin / Q000235:genetics
D002021:Buffers; D006146:Guanidines; D017879:Isothiocyanates; D012333:RNA, Messenger; D011989:Receptors, Thyrotropin; D012996:Solutions; D001613:beta 2-Microglobulin; C054435:guanidine isothiocyanate; D004247:DNA; D013954:Thyroglobulin