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An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
Published on: January 11, 2017
A minimum requirements method to isolate large quantities of highly purified DNA from one drop of poultry blood
1Department of Animal Production, College of Agriculture, Al-Qasim Green University, 8-Al Qasim, 51001 Babil, Iraq. mohammed79@agre.uoqasim.edu.iq, baquralhilly_79@yahoo.com.
Abstract:
Here, we describe a rapid, efficient and noncomplicated method for extracting poultry genomic DNA. This simple method obtains excellent qualities of molecular biology grade DNA in a matter of only about 15 min. A straightforward protocol is followed in this extraction procedure, in which, when the blood cells are placed in a distilled water alone, water rapidly disrupts red blood cells (RBCs) membranes and alleviate chickens' undesired high blood viscosity without being aided by any other chemicals. Moreover, the time, cost and efforts were hugely reduced since the step of leukocytes lysis was successfully mixed with the protein precipitation. The isolated genomic DNA, in terms of its quantity and quality, is satisfying and is proved to be suitable for restriction endonucleases digestion, polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLP). Thus, instead of extracting a limited amount of DNA using expensive or relatively expensive kits, this guaranteed procedure can be utilized alternatively to accurately extract genomic DNA (gDNA) from minimal starter quantities of the chicken's blood that do not exceed 50 μL. By relying on this technique, all the practical handover steps are reduced to the extreme limits. This pilot study may provide a high yield gDNA extraction that minimizes labour, expense, and steps in very high competency and reproducibility.
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