Optimization of DNA extraction and amplification of Kikuyu (Pennisetum clandestinum Hochst. ex Chiov) for molecular identification

Kikuyu (Pennisetum clandestinum Hochst. ex Chiov) is an important forage containing high crude protein for livestock. Molecular analysis of kikuyu relies on high yields of pure DNA and suitable PCR conditions. This research aimed to extract DNA from kikuyu based on weight of the sample and amplify the DNA of Burangrang accession using specific primers. 100 grams and 200 grams leaves of 3 accessions of kikuyu from Burangrang, Bukit Tunggul, and Tangkuban Perahu were extracted by Qiagen Mini Kit Plant. Concentration and purity of DNA were analyzed by NanoDrop Spectrophotometer 2000. DNA from Burangrang accession was amplified using six specific primers at different annealing temperatures. The result showed that the yield of DNA ranged 2.2 µg/µl to 21.4 µg/µl and the purity (ratio) were 1.08 to 2.01. Bukit Tunggul and Burangrang accession showed the same interaction pattern on the sample weight for concentration and purity. One hundred grams of leaves from Burangrang accession produce the highest concentration and the best purity of DNA, but no difference between other weight and accession. Reproducible amplifiable products were observed in all PCR reactions except primer K2. These results indicated that optimized protocol is suitable for further work on molecular identification of kikuyu.