
A Simple, Efficient and Rapid Method for Good Quality DNA Extraction from Rice Grains
Received date: 2016-08-02
Accepted date: 2016-09-11
Online published: 2016-12-28
An efficient and good DNA extraction protocol should be simple, affordable and yield enough DNA with high quality. Rice (Oryza sativa L.) DNA extraction methods often use seedlings or leaves rather than the grains and tend to be time-consuming, involve multiple steps, and use hazardous chemicals and expensive enzymes. Rice grains offer several benefits over seedlings and leaves as a source of DNA for genetic analysis. However, these benefits are underutilized because the bulk of a rice grain is made up of starch. It is particularly important, but difficult to get rid of the starch while extracting DNA from rice grains. This co-precipitated polysaccharide is a known inhibitor of DNA polymerase activity in polymerase chain reaction (PCR). We describe here a very simple and highly affordable Chelex®-100 based DNA extraction method from rice grains. It does not require any hazardous chemicals or enzymes. This method reproducibly extracts DNA with good purity indices (A260/A230 and A260/A280 values), but requires only a few steps.
Key words: DNA extraction; rice; grain; Chelex®-100
Ashfaqur Sajib Abu, Ashraful Islam Bhuiya Mohammad, Huque Roksana . A Simple, Efficient and Rapid Method for Good Quality DNA Extraction from Rice Grains[J]. Rice Science, 2017 , 24(2) : 119 -122 . DOI: 10.1016/j.rsci.2016.09.003
| 1 | Ahmadikhah A.2009. A rapid mini-prep DNA extraction method in rice (Oryza sativa).Afr J Biotechnol, 8(2): 323-328. |
| 2 | Aliyu R E, Wasiu A A, Dangora D B, Ademola J O, Adamu A K.2013. Comparative study of genomic DNA extraction protocols in rice species.Int J Adv Res Technol, 2(2): 1-3. |
| 3 | Beyenal H, Babauta J T.2015. Biofilms in Bioelectrochemical Systems: From Laboratory Practice to Data Interpretation. Hoboken, USA: John Wiley & Sons. |
| 4 | Butler J M.2005. Forensic DNA Typing: Biology, Technology, and Genetics of STR Markers. 2nd edn. London, UK: Elsevier Academic Press. |
| 5 | Casquet J, Thebaud C, Gillespie R G.2012. Chelex without boiling, a rapid and easy technique to obtain stable amplifiable DNA from small amounts of ethanol-stored spiders.Mol Ecol Resour, 12(1): 136-141. |
| 6 | Chen W Y, Cui H R, Bao J S, Zhou X S, Shu Q Y.2006. A simplified rice DNA extraction protocol for PCR analysis.Rice Sci, 13(1): 67-70. |
| 7 | Chunwongse J, Martin G B, Tanksley S D.1993. Pre-germination genotypic screening using PCR amplification of half-seeds.Theor Appl Genet, 86(6): 694-698. |
| 8 | Duan Y B, Zhao F L, Chen H D, Li H, Ni D H, Wei P C, Sheng W, Teng J T, Zhang A M, Xue J P.2015. A simplified genomic DNA extraction protocol for pre-germination genotyping in rice.Genet Mol Res, 14(2): 6369-6375. |
| 9 | Evamoni F Z, Rubel M H, Hossain M A.2014. Genetic variation and relatedness for blb resistance in BRRI rice using RAPD markers.Int J Innov Appl Stud, 8(1): 93-106. |
| 10 | Ferdous J, Hanafi M M, Rafii M Y, Muhammad K.2012. A quick DNA extraction protocol: Without liquid nitrogen in ambient temperature.Afr J Biotechnol, 11(27): 6956-6964. |
| 11 | Fouladvand A, Yari H, Emami A, Khosravi H R M, Pourmehdi S.2013. Optimization of a rapid DNA extraction protocol in rice focusing on age of plant and EDTA concentration.J Med Bioengin, 2(3): 218-223. |
| 12 | Ibemhal D A, Majumder P B, Imotomba R K, Laishram J M.2015. Genomic DNA extraction from the black scented rice (Chakhao).Ind J Plant Sci, 4(1): 34-37. |
| 13 | Kanawapee N, Sanitchon J, Srihaban P, Theerakulpisut P.2011. Genetic diversity analysis of rice cultivars (Oryza sativa L.) differing in salinity tolerance based on RAPD and SSR markers.Electr J Biotechnol, 14(6): 1-17. |
| 14 | Liang H B, Deng Y, Wang C T, Xu X.2015. A high-throughput DNA extraction method from rice seeds.Biotechnol Biotechnolog Equip, 30(1): 32-35. |
| 15 | Mutou C, Tanaka K, Ishikawa R.2014. DNA extraction from rice endosperm (including a protocol for extraction of DNA from ancient seed samples).Methods Mol Biol, 1099: 7-15. |
| 16 | Roychowdhury R, Karmakar J, Dey N.2012. PCR-compatible genomic DNA isolation from different tissues of rice (Oryza sativa) for SSR fingerprinting.Eur Asian J BioSci, 6: 85-90. |
| 17 | Sun C, He Y H, Chen G, Rao Y C, Zhang G H, Gao Z Y, Liu J, Ju P N, Hu J, Guo L B, Qian Q, Zeng D L.2010. A simple method for preparation of rice genomic DNA.Rice Sci, 17(4): 326-329. |
| 18 | Thomas J, Dominic V J.2015. Isolation of high quality DNA from pulverized rice grain samples by tri-phase method.Int J Sci Res, 4(11): 524-527. |
| 19 | Vibhuti M J, Viralkumar B M, Vrinda S T.2015. Simple and efficient protocol for RNA and DNA extraction from rice (Oryza sativa L.) for downstream applications.Int Res J Biol Sci, 4(2): 62-67. |
| 20 | Walsh P S, Metzger D A, Higuchi R.1991. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material.Biol Technol, 10(4): 506-513. |
/
| 〈 |
|
〉 |