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Characterization of Different Types of Native Sheep by Using Modern Techniques of Breeding and Genetics.

M. N. Hasan
Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

S. Pervage
Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka1341.

M. Ershaduzzaman
Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

M. A. I Talukder
Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

M. A. M. Y. Khandoker
Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202.

Abstract/ Executive Summary:

Blood samples of three regional sheep breeds of both sexes were collected from the farm of the Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute (BLRI), Savar, Dhaka 1341. Whole blood was collected from the jugular vein into 8 ml vacutainer tubes containing the EDTA anticoagulant. Blood samples were kept cold at 4?. DNA was extracted by equal volumes of phenol, phenol-chloroform (1:1) and chloroform-isoamylalcohol 24:1. DNA was precipitated by adding 2 volumes of chilled ethanol in the presence of a high concentration of salts (10% 3 M sodium acetate). The pellet was washed with 70% ethanol, air-dried and subsequently dissolved in TE buffer (10 mM Tris-HCI, 1 mM EDTA). The PCR reactions (25µl) contained 1 U Taq DNA polymerase; 2.5 µl dNTPs; 2.5 V Tris buffer (pH=8.8; 1.5 µl MgC12; 10.3 µl dH2O; 2.5 µl glycerin; 50 ng of random primer and 20-100 ng of DNA. ? (1 min), 36? (1 min) and 72? (2 min) with an additional extension step for 10 min. Amplification products were visualized by electrophoresis in 1.4 % agarose gels in 1x TBE buffer in the presence of ethidium bromide. The gels were photographed under UV light. The amplified DNA fragments were not found. The research work would be continued using RAPD marker to estimates the genetic distances or similarities among different types of native sheep in Bangladesh.

Keywords: Characterization, Native sheep, Modern techniques

Location:

Start Date: 01-07- 2009

End Date: 30-06-2010

Program Area: Variety and Species

Commodity: Sheep

Objectives:

Find out the status of amplified DNA fragments for characterization of different types of native sheep by using modern techniques of Breeding and Genetics.

Methodology:

The protocol of the current study may explain in the following: Collection of blood, Extraction of DNA, Quantification of DNA, Random Amplification of polymorphic DNA (RAPD). Blood samples of three regional sheep breeds of both sexes were collected from the farm of the Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute (BLRI), Savar, Dhaka 1341. Whole blood was collected from the jugular vein into 8 ml vacutainer tubes containing the EDTA anticoagulant. Blood samples were kept cold at 4?. DNA extraction was carried out as fellows: to an aliquot of 100 µl blood (after thawing), 700 µl of lyses buffer (10 mM Tris-HCI, 100 mM NaC1, 1 mM EDTA, pH 8.0, .5% SDS) and 60 µg of proteinase K (20 mg/ml) were added, The mixture was vortexed and incubated at 37? overnight. DNA was extracted by equal volumes of phenol, phenol-chloroform (1:1) and chloroform-isoamylalcohol 24:1. DNA was precipitated by adding 2 volumes of chilled ethanol in the presence of a high concentration of salts (10% 3 M sodium acetate). The pellet was washed with 70% ethanol, air-dried and subsequently dissolved in TE buffer (10 mM Tris-HCI, 1 mM EDTA). All oligonucleotide primers were 10 to 16 – per. Forthy of this primers were reported by Kantanen et al. (1995) that can produce repeatable bands in sheep and remainder three primers will be chosen according to unpublished data (Table 1). The PCR reactions (25µl) contained 1 U Taq DNA polymerase; 2.5 µl dNTPs; 2.5 V Tris buffer (pH=8.8; 1.5 µl MgC12; 10.3 µl dH2O; 2.5 µl glycerin; 50 ng of random primer and 20-100 ng of DNA. Following an initial step at 94? for 2 min, the reaction were subjected to 35 cycles of Amplification at 94? (1 min), 36 ? and 72? (2 min) with an additional extension step for 10 min. Amplification products were visualized by electrophoresis in 1.4 % agarose gels in 1x TBE buffer in the presence of ethidium bromide. The gels were photographed under UV light.

Source of Information: BLRI. Proceedings of the Annual Research Review Workshop 2009-2010, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The amplified DNA fragments were not found. The research work is repeating and expecting using RAPD marker with PCR kit to estimates the genetic distances among different types of native sheep in Bangladesh.

Knowledge Management: Report/Proceedings