Genomic DNA Extraction from Different Human Tissue Sample: Urine, Oral Gargle, Blood, and Cervical for Real Time Amplification by qPCR

AbstractMolecular technologies have been a driver of rapid paradigm shift in the field of scientific research and clinical applications. Adequate and pure form of nucleic acid extracted from efficient biological sources together with an easy, simple, standardized protocol, ease of access and acceptability is also a prerequisite for genotyping analysis for different disorders. Urine and oral gargle samples are emerging as a potential source of genomic DNA (gDNA) and may offer a better alternative to existing sources such as blood. This manuscript compares for the first time, the quality and quantity of gDNA extracted manually by standard methods from different biological samples like urine, oral gargle, blood and cervical samples, together. It aimed to assess the feasibility of extracting sufficient gDNA from easily accessible and acceptable samples for amplification. 646 urine, oral, blood and cervical samples were collected from different studies in the department and analyzed. gDNA from blood was extracted by Proteinase K digestion followed by ethanol precipitation, and phenol –chloroform extraction method for urine, oral gargle and cervical specimen. The quantification of isolated gDNA were analyzed in Nano-drop spectrophotometer and 1% Agarose gel electrophoresis for different sample types. Humanβ-globin gene were used for internal quality control for the real time amplification. DNA quantity and purity were adequate and comparable for amplification in all the three bi...
Source: Indian Journal of Clinical Biochemistry - Category: Biochemistry Source Type: research