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Updated: Sep 1, 2025
Collection and Extraction of Saliva DNA for Next Generation Sequencing
Published on: August 27, 2014
Rapid and inexpensive method of PCR ready DNA isolation from human peripheral blood and saliva
Rajat Hegde1, Smita Hegde1, Pooja Joshi1
1Karnataka Institute for DNA Research (KIDNAR), Dharwad, 580003, Karnataka, India.
Background:
The isolation of nucleic acids is a frequently performed procedure in the molecular biology area. Although several rapid DNA isolation techniques from human peripheral blood and saliva have been developed, there are still some disadvantages - volume, time, cost, and yield are a few notable ones.
Objective:
We aim to develop a rapid and inexpensive method to isolate high-molecular-weight genomic DNA from human peripheral blood and saliva that can be used for molecular biology experiments.
Methods:
Five DNA isolation methods with slightly varying protocols were used. High-quality DNA obtained from one specific method was further amplified by PCR and the template with good amplification was further used for performing RFLP and sequencing.
Results:
Out of 5 different isolation methods (R1 to R5), DNA obtained from the R4 was of good quality (molecular weight is > 10 kb and 260/280 ratio is 1.89 ± 0.2), which allows successful PCR amplification and good separation in Restriction Fragment Length Polymorphism analysis. Sequencing by the Sanger Sequencing produced a good readable sequence of an amplified fragment from Method R4 DNA.
Conclusion:
In the present study we have developed a simple, rapid, and cost-effective DNA isolation method, which uses low sample volume and yields good quantity and high-quality product. The DNA obtained is highly fit for molecular genetics research applications.
