A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples

Abstract

Background Different methods available for extraction of human genomic DNA suffer from one or more drawbacks including low yield, compromised quality, cost, time consumption, use of toxic organic solvents, and many more. Herein, we aimed to develop a method to extract DNA from 500 μL of fresh or frozen human blood. Methods Five hundred microliters of fresh and frozen human blood samples were used for standardization of the extraction procedure. Absorbance at 260 and 280 nm, respectively, (A260/A280) were estimated to check the quality and quantity of the extracted DNA sample. Qualitative assessment of the extracted DNA was checked by Polymerase Chain reaction and double digestion of the DNA sample. Results Our protocol resulted in average yield of 22±2.97 μg and 20.5±3.97 μg from 500 μL of fresh and frozen blood, respectively, which were comparable to many reference protocols and kits. Conclusion Besides yielding bulk amount of DNA, our protocol is rapid, economical, and avoids toxic organic solvents such as Phenol. Due to unaffected quality, the DNA is suitable for downstream applications. The protocol may also be useful for pursuing basic molecular researches in laboratories having limited funds.

A Novel DNA Extraction Protocol from Frozen Blood of Normal Individuals and Patients Who Received Systemic Chemotherapy<strong> </strong>

“…This piece of evidencein turnis very beneficial in terms of reducing costs and efforts without scarifying DNA in these samples. Add to that, the non-organic / non-enzymatic steps have also proven more feasibility of our protocol as it does not require the hazardous inclusion of toxic organic solvents [13], or the extended incubation with proteinase K [25]. However, the utilization of this protocol is not restrained with these chemotherapeutic cases, it can be used to extract DNA with a considerable efficiency for downstream molecular biology experiments.…”

“…With the progressively prescribed chemical treatments of tumors, the negative impact of these drugs on the extracted DNA is increasingly being confronted. However, in addition to the commonly known methods [9,10,11], series of DNA extraction protocols have recently been devised for DNA extraction from blood [12,13,14]. Despite the immense progress made in DNA extraction, little attention has been paid to reduce the difficulty of undergoing downstream experiments for DNA samples extracted from patients who are exposed to regular chemotherapeutic sessions.…”

The correlation between the level of 3-hydroxypropyl mercapturic acid, CYP2B6 polymorphisms, and hematuria occurrences after cyclophosphamide administration and its bioanalytical methods: A systematic review

“…47 Blood sample is better from the other biological samples because it contains nucleated white blood cells and there are many protocols regarding DNA isolation from blood that J o u r n a l P r e -p r o o f have been published. 48 Therefore, we conclude that it is best to use blood samples to carry out a DNA extraction process.…”

“…Of various biological samples, blood got the most important attention in biological research. 48 However, blood is the most complex biological sample because many components in it can interfere with the process of bioanalysis. Handling of blood samples also needs to be careful because too much water can cause blood cells to undergo hemolysis.…”

Effects of an APOE Promoter Polymorphism on Fronto-Parietal Functional Connectivity During Nondemented Aging

“…Genomic DNA was extracted from the blood samples of the subjects using the standard method (Guha et al, 2018 ). The participants were pre-screened for APOE genotype using a TaqMan single nucleotide polymorphism (SNP) genotyping assay on a 7900 HT Fast Real-Time PCR system (Applied Biosystems, Foster City, CA, USA; Seripa et al, 2011 ).…”