A simple and efficient protocol for isolation of genomic DNA from shrimps’ old tissues of Green Tiger Shrimp,Penaeus semisulcatus: Suitable for Inverse PCR

Document Type : Genetic

Authors

1 Department of Marine Biotechnology, School of Marine Science and Technology, University of Persian Gulf, Bushehr

2 Department of Biotechnology, Institute of Persian Gulf, Persian Gulf University, Bushehr

3 Department of Cell and Molecular Biology, Faculty of Science, University of Persian Gulf, Bushehr

Abstract

The selection of an appropriate protocol for intact DNA extraction from shrimps’  old tissues that have been preserved in alcohol for a long time  has an important role in the success of molecular biology analysis. The aim of current study was to optimize a simple, reproducible, safe and reliable protocol for quality genomic DNA extraction from old muscular tissues of Green Tiger Shrimp, Penaeus semisulcatus. The main advantages of this procedure involved Pulverizing the tissue in a mortar after being dried in  40 ° C Temperature,   avoiding the use of  liquid nitrogen for tissue pulverization  and the reducing ot extraction time to 5 hr. Because the success Inverse PCRis highly dependent on the quality of DNA temolate, the isolation of a part of beta actine gene using this reaction for the evaluation of the quality of the extracted DNA was considered.  Success of amplification and sequencing of the beta actin gene of the Green Tiger Shrimp indicated that the resulting DNA from this procedure had necessary for this reaction. Regarding the success of the Invers PCR  the resulting DNA from this protocol can be successfully used as a template in other molecular studies such as AFLP, RFLP and the amplification of long fragments which require quality DNA.

Keywords

Journal of Animal Environment
Volume 8, Issue 4
January 2017
Pages 227-236

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