Extracting method for sugarcane chloroplast DNA and its verification

【Objective】A lab-friendly extracting method for sugarcane cpDNA was explored based on sugarcane traits in order to provide references for molecular biology research of cpDNA.【Method】Referring to DNase I differential centrifugation treatment of rice and wheat cpDNA,sugarcane cpDNA was extracted and purified using the modified procedure.trnL gene sequencing was compared and analyzed to verify the feasibility of cpDNA extraction.【Result】 High quality(OD260/OD280:1.79-1.90) and high amount of cpDNA(2.71 μg per gram of fresh leaves) was isolated and detected by Eppendorf tester of protein nucleic acid.The trnL genes in extracted cpDNA were comparatively analyzed.The results showed that trnL genes of the tested species(cultispecies,Saccharum spontaneum hybrid and backcross) were 99% homologous with the reported cultivated species NCo 310,SP-80-3280 and Saccharum spontaneum HN0046.Base mutation,insertion and deficiency occurred in eight loci like 381,386,389,392,393,400,488,490 bp.【Conclusion】This modified method can serve as an efficient cpDNA extraction procedure for further molecular research.