Isolation of genomic DNA from Sea Cucumber /
Maria Cristina Denate
- 2001
- 64 leaves
Thesis (BS Biology) -- University of the Philippines Mindanao, 2001
Isolation of genomic DNA from muscle of sea cucumber was done in order to compare the different species collected from Davao Gulf. The extraction was attempted using two techniques. In the first procedure, muscle was frozen in liquid nitrogen, homogenized, and the DNA extracted using phenol-chloroform and alcohol precipitation. In the second procedure, extraction was done using a commercial kit DNAzol (Gibco) and further precipitated with ethanol. PCR amplification using 2 primer pairs (random hexamers and actin gene) was done to evaluate genomic DNA integrity. Random hexamers did not show any band while human beta actin gene showed a 318bp band in two out of five species. This band was further restricted using 3 enzymes. Hinf1, Mval, and Scrf1. The use of these 3 restriction enzymes is a promising method of studying the genetics of Holothuria. While this attempt shows that the DNAzol extraction appears to be a more efficient method, it leaves several questions for further investigation