DNA profiling of Blumea balsamifera (L.) DC. using direct amplification of length polymorphisms (DALP) analysis

• Main Author: Chin, Joshua.
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak (UNIMAS) 2006
• Subjects: QR Microbiology
• Online Access: http://ir.unimas.my/id/eprint/18225/8/DNA%20profiling%20of%20Blumea%20balsamifera%20%28L.%29%20DC.%20using%20direct%20amplification%20of%20length%20polymorphisms%20%28DALP%29%20analysis%20%28fulltext%29.pdf; http://ir.unimas.my/id/eprint/18225/
• Institution: Universiti Malaysia Sarawak

Studying DNA polymorphism in medicinal plant such as Blumea balsamifera is valuable for genetic manipulation such as trait improvement. There is a need to establish a suitable genomic DNA extraction protocol, as the plant is high in phenolic compounds, which can inhibit PCR reaction. Thus, three extraction methods were carried out Subsequently, the DNA extraction method using high PVP percentage (6 %) was detennined to be the most effective, yielding intact DNA of high quality, with insignificant levels of contaminations. Using that method, DNA from four locations were extracted and then subjected to PCR amplification using DALP analysis to generate DNA profiles. The selective forward primer DALP 221, paired with DALPR reverse primer successfully generated multiple banding patterns. Ten different DNA bands were generated ranging from 150 bp to 1500 bp and the DALP profile was reproducible. Four constant bands were observed in all the locations. DNA samples from Asa Jaya and Bau have the same banding patterns. DNA samples from Samarahan and Kuching have different banding patterns compared to those from Asa Jaya and Bau. Three bands were absent in Samarahan's DNA sample but present in the other locations. In tead, two different bands, not observed in other locations, were present in DNA sample from Samarahan. DNA sample from Kuching has the same seven of the eight bands that were shared among DNA samples from Asa Jaya and Bau. However, there was an absence of a band observed in Kuching's DNA sample. Thus, Samarahan and Kuching exhibit certain degree of DNA variations. However, further verification through statistical analysis is needed to confirm DNA polymorphism in future studies.