Experience of quantity and quality of DNA and RNA extraction from limited pediatric blood samples: A comparative analysis of automated and manual kit-based method

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dc.contributor.authorSharma, Pen_US
dc.contributor.authorSingh, Men_US
dc.contributor.authorSingh, Aen_US
dc.contributor.authorBhardwaj, Den_US
dc.contributor.authorBhatia, P.en_US
dc.date.accessioned2023-08-10T07:32:13Z
dc.date.available2023-08-10T07:32:13Z
dc.date.issued2022-03
dc.description.abstractIntroduction: Optimal DNA and RNA quantity and purity is essential for downstream molecular biology experimentation and to avoid re-processing of sample. Despite availability of different kits and automated systems for nucleic acid isolation there is limited data on their performance evaluation, more so with pediatric blood samples, that are usually compromised in quantity. Hence, we evaluated the performance of automated QIAcube platform using pediatric blood samples in parallel with manual Qiagen extraction kits. Materials and Methods: A total of 500 samples were analyzed based on groups of PBMC and direct blood input. The isolated DNA and RNA were surveyed for quantity and quality tests by spectrophotometric and downstream analysis. Results: There was no significant difference in the DNA quantity (ng/ul) between manual and automated method based on similar sample input but quality (260/280) was significantly better with the QIAcube platform when direct blood and or PBMCs were used for extraction respectively (1.82 ± 004 Vs. 1.84.002; P-0.000008 and 1.859 ± 005 Vs. 1.843 ± 0.003; P-0.02). Moreover, the standard error mean was low for both quantity and quality in the QIAcube method suggesting uniformity. Comparison of quality assessment by spectrophotometer and qubit fluorimeter showed that QIAcube sheared DNA less (P- 0.038) as compared to manual method (P-0.013). Also, time taken to process the samples in QIAcube was 23% less than the kit-based method. Conclusion: Overall analysis of QIAcube platform suggests that it yields more better, uniform, and less-sheared quality of nucleic acid in a relatively less time as compared to manual extraction kits.en_US
dc.identifier.affiliationsPediatric Hematology-Oncology Laboratory, Advanced Pediatric Centre, Postgraduate Institute of Medical Education and Research, Chandigarh, Indiaen_US
dc.identifier.citationSharma P, Singh M, Singh A, Bhardwaj D, Bhatia P.. Experience of quantity and quality of DNA and RNA extraction from limited pediatric blood samples: A comparative analysis of automated and manual kit-based method. Indian Journal of Pathology & Microbiology. 2022 Mar; 65(1): 105-110en_US
dc.identifier.issn0377-4929
dc.identifier.issn0974-5130
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/223178
dc.languageenen_US
dc.publisherWolters Kluwer - Medknowen_US
dc.relation.issuenumber1en_US
dc.relation.volume65en_US
dc.source.urihttps://doi.org/10.4103/IJPM.IJPM_946_20en_US
dc.subjectDNAen_US
dc.subjectkitsen_US
dc.subjectqiacubeen_US
dc.subjectqualityen_US
dc.subjectquantityen_US
dc.subjectRNAen_US
dc.titleExperience of quantity and quality of DNA and RNA extraction from limited pediatric blood samples: A comparative analysis of automated and manual kit-based methoden_US
dc.typeJournal Articleen_US
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