Quality of Cold Centrifuge Affect on PCR Results
Keywords:
HLAB-27, DNA Extraction, PCR.
Abstract
See Full text DNA extraction is an important step for PCR test. We are doing DNA extraction by salt-out and Phenol Chloroform method using two cold centrifuge instruments, one PSN Instrumentation Pvt. Ltd, Delhi Company and the other one Plasto Craft industries (P) Ltd. Mumbai. We performed HLAB-27 typing by conventional allele-specific PCR and the kit of BAG Company supplied by M/S Shiva Scientific. Using these two methods, we found that DNA extracted in PSN cold centrifuge gave false positive band in most cases while DNA extracted by Plasto Craft cold centrifuge did not give any false positive result in HLAB-27. Up till now it was considered that type and amount of sample, its collection, pipettting, quality of distilled water, contamination and quality of reagents, thermal cycle programme and gel electrophoresis affect PCR results. However our study suggest that quality of cold centrifuge also affect the quantity of DNA, PCR results and give false positive results and hence cold centrifuge should be of good quality and calibrated.References
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2. Bon MAM, Oeveren-Dybicz AV, Bergh FAJTMVD. Genotyping of HLA-B27 by Real-Time PCR without Hybridization Probes, Clinical Chemistry, Technical Briefs 2000; 46(7): 1000-02.
3. Michael N, Andreas B, Christoph W. Fundamentals of quality assessment of molecular amplification methods in clinical diagnostics. Clinical Chemistry1998; 44(1):12–26.
4. Jaber A. Brief Review:Preventing PCR Amplification Carryover Contamination in a Clinical Laboratory. Annals of Clinical & Laboratory Science 2004; 34(4):389-96.
5. Rys PN, Persing DH. Preventing False Positives: Quantitative Evaluation Of Three Protocols For Inactivation Of Polymerase Chain Reaction Amplification Products, Journal Of Clinical Microbiology 1993; September: 2356-60.
Published
2015-10-04
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Section
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