OPTIMIZATION OF DNA EXTRACTION PROTOCOLS FROM SHRIMP (Penaeus monodon) TISSUE

B.  Biswas; M.N.  Islam; M.N.  Siddiqui; M.N.  Ahsan

doi:10.53808/KUS.2008.9.2.0817-L

Authors

B. Biswas Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh
M.N. Islam Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh
M.N. Siddiqui Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh
M.N. Ahsan Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

DOI:

https://doi.org/10.53808/KUS.2008.9.2.0817-L

Keywords:

DNA extraction, DNA concentration, extraction protocols, optimization.

Abstract

DNA analyses and DNA-based diagnoses are going forwardly in the commercial deals with shrimp (Penaeus monodon) as well as in the basic research. In this regard extraction of DNA is obvious. However, numerous protocols are available DNA extraction from animal tissue. Therefore, which technique is suitable one for shrimp tissue is indispensable to justify. In the current study, a protocol was optimized for extraction of DNA from shrimp collected from Batiaghata, Khulna. Muscle Tissue (MT) and Hepatopancreas Tissue (HT) were distinctly sampled on15^th March, 2007 from only one fresh individual for each of six protocols applied. Guanidium isothiocyanate treatment involving two groups of commercial preparation (DNAzol from invitrogen^® and DNA Extraction Buffer (DEB) from Genei^®) with or without RNAse, as well as Proteinase K isolation technique and sodium dodecyle sulfate (SDS) isolation technique were applied to extract DNA. The quantity (µgml^-1) and quality were determined by UV spectrophotometry at A_260nm and A_260/280 respectively. The current study provided that the quantity of DNA obtained with almost every protocol was higher when the hepatopancreas tissue was used. Among six protocols, DEB with RNAse (102.5 µgml^-1 for MT, 302.5 µgml^-1 for HT) resulted the highest concentration of DNA with acceptable level of purity (A_260/280 1.86 for MT and 1.70 for HT).

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