Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

Extraction of Jute Genomic DNA: Difficulties and Solutions

A. K. M. Shahadat Hossain
SSO
Jute Agricultural Experimental StationJaigir, Manikganj.

Md. Abdul Latif
MS Student
Department of Biotechnology, BAU

Bipul Kumar Biswas
MS Student
Department of Biotechnology, BAU

Saidin Saclain
MS Student
Department of Biotechnology, BAU

Md Salahuddin
MS Student
Department of Physiology, BAU

M. Abu Sayed
MS Student
Department of Biochemistry and Molecular Biology, HMDSTU

Md. Mahboob Hussain
PSO
Genetic resources and Seed Division BJRI, Dhaka-1207.

Sheik Md Moniruzzaman
MS Student
Department of Biochemistry and Molecular Biology, BAU

Md. Shahidul Islam
Associate Professor
Department of Biotechnology BAU, Mymensingh, Bangladesh.

Abstract/ Executive Summary:

The plant parts of jute are composed of highly viscous substances and phenolic compounds which make nuisance to extract good quality genomic DNA (gDNA). Here different methods viz., phenol chloroform isoamylalcohol extraction and ethanol precipitation method; potassium acetate method and Cetyl trimethylammonium bromide (CTAB) method were applied for isolating the jute gDNA from leaves of 14 and 40 day’s plants. All of these methods were unable to extract good quality gDNA from jute leaves. That is why, in this experiment, the concentration and chemical compositions of CTAB method were modified for obtaining good quality jute gDNA. Polyvinylpyrrolidone (PVP) was added in the CTAB extraction buffer and β mercaptoethanol was used while grinding the leaf tissues with CTAB extraction buffer. It was found that good quality gDNA was obtained using modified CTAB method from 14 day’s plant’s leaves but low quality gDNA was obtained from 40 day’s plant’s leaves. These were confirmed by 0.8% agarose gel electrophoresis. Considering the visual quality of the banding patterns and their reproducibly MHR24 (5/-TTCCCTCCCA-3/) and MHR21 (5-/CCCGAAGCGA-3/) primers were selected out of 8 Random Amplified Polymorphic DNA (RAPD) primers for PCR. A total number of 11 loci were identified by these RAPD primers. Popgen32 software was used for analyzing the RAPD data. The number of polymorphic loci was one and the percentage was 9.09 which stated that a low level of genetic variations was existed among the jute accessions. In the dendrogram, jute accession CC875 (C. capsularis) was grouped in one cluster while CC894 (C. capsularis) and CC896 (C. capsularis) accessions were grouped in another cluster.

Keywords: Jute; DNA extraction; Phenolic compounds; RAPD.

Location: Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Start Date: 05-10-2013

End Date: 23-12-2013

Program Area: Variety and Species

Commodity: Jute

Objectives:

The main objective was to overcome the problems associated with jute gDNA extraction by modifying CTAB method.

Methodology:

Potassium acetate method, phenol chloroform isoamylalcohol extraction and ethanol precipitation method, CTAB method were used to extract jute gDNA but good quality gDNA was not obtained using these methods. Finally, CTAB method was modified for obtaining good quality gDNA. The main activities were:

Composition of CTAB extraction buffer and preparation
Leaf collection and preparation
Extraction and purification of gDNA
Removal of RNA
Quantification of DNA (Spectrophotometric method)
PCR conditions and data analysis

Source of Information: Int J Appl Sci Biotechnol, Vol 2(4): 516-520, 2014

Article Link (Online Journal):

Funding Source:

1. Government

Budget: 50,000

Total Budget (Tk.) : 50,000

Achievement/Findings:

To overcome the problems associated with jute gDNA extraction of, CTAB method has been modified.

Knowledge Management: Journal