| Freezing complete polymerase chain reaction master mix reagents for routine molecular diagnostics. | |
| | |
MedLine Citation:
|
PMID: 17121086 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The potential of storing complete frozen real-time polymerase chain reactions (PCRs) and real-time reverse transcription PCRs (RT-PCRs), which require only thawing and the addition of template nucleic acid before PCR cycling, was examined. Master mixes containing all necessary reagents at working concentration (except template nucleic acid) were aliquoted into single-reaction volumes and stored at -70 degrees C for periods of up to 8 months. Reactions were removed from storage and nucleic acid template was added and amplified using different real-time PCR instruments. Threshold cycle values were used to monitor changes in assay performance after storage. Results for hybridization probe and TaqMan probe assays showed that freezing complete real-time PCR and RT-PCR reaction mixes was possible without deterioration in assay performance. This approach has advantages for routine molecular diagnostics in areas such as convenience, test consistency, quality control, and ease of use by nonspecialist staff. |
| | |
Authors:
|
Donna Marie West; Jason Sawyer |
Related Documents
:
|
14620816 - Soluble sunscreens fully protect e. coli from disinfection by electrohydraulic discharges. 4975366 - Physical and chemical properties of the transmissible mink encephalopathy agent. 22578196 - Orally administered [14c]dpa and [14c]dha are metabolised differently to [14c]epa in rats. 7721866 - A yeast gene product, g4p2, with a specific affinity for quadruplex nucleic acids. 20005536 - Serum uric acid and metabolic syndrome in taiwanese adults. 20359186 - Gastroprotective effect of carnosic acid gamma-lactone derivatives. |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc Volume: 18 ISSN: 1040-6387 ISO Abbreviation: J. Vet. Diagn. Invest. Publication Date: 2006 Nov |
Date Detail:
|
Created Date: 2006-11-22 Completed Date: 2008-05-06 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 9011490 Medline TA: J Vet Diagn Invest Country: United States |
Other Details:
|
Languages: eng Pagination: 580-2 Citation Subset: IM |
Affiliation:
|
Technology Transfer Unit, Biotechnology Department, Veterinary Laboratories Agency-Weybridge, New Haw, Addlestone, Surrey, United Kingdom. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Cattle Cryopreservation* DNA Probes / standards* DNA, Bacterial / analysis DNA, Mitochondrial / analysis Deoxyribonucleases / diagnostic use Leptospira / genetics Mammals Polymerase Chain Reaction / methods*, standards RNA, Viral / analysis Reverse Transcriptase Polymerase Chain Reaction / methods, standards Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
|
0/DNA Probes; 0/DNA, Bacterial; 0/DNA, Mitochondrial; 0/RNA, Viral; EC 3.1.-/Deoxyribonucleases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Aplasia cutis congenita (epitheliogenesis imperfecta) in swine: observations from a large breeding h...
Next Document: Comparison of direct fluorescent antibody staining and real-time polymerase chain reaction for the d...