A simplified universal genomic DNA extraction protocol suitable for PCR.

نویسندگان

  • T Y Wang
  • L Wang
  • J H Zhang
  • W H Dong
چکیده

Conventional genomic DNA extraction protocols need expensive and hazardous reagents for decontamination of phenolic compounds from the extracts and are only suited for certain types of tissue. We developed a simple, time-saving and cost-efficient method for genomic DNA extraction from various types of organisms, using relatively innocuous reagents. The protocol employs a single purification step to remove contaminating compounds, using a silica column and a non-hazardous buffer, and a chaotropic-detergent lysing solution that hydrolyzes RNA and allows the selective precipitation of DNA from cell lysates. We used this system to extract genomic DNA from different tissues of various organisms, including algae (Dunaliella salina), human peripheral blood, mouse liver, Escherichia coli, and Chinese hamster ovary cells. Mean DNA yields were 20-30 μg/cm(3) from fresh tissues (comparable to yields given by commercial extraction kits), and the 260/280 nm absorbance ratio was 1.8-2.0, demonstrating a good degree of purity. The extracted DNA was successfully used in PCR, restriction enzyme digestion and for recombinant selection studies.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Rapid DNA Extraction Protocol from Stool, Suitable for Molecular Genetic Diagnosis of Colon Cancer

Colorectal cancer (CRC) is one of the most common forms of cancers in the world and is curable if diagnosed at the early stage. Analysis of DNA extracted from stool specimens is a recent advantage to cancer diagnostics. Many protocols have been recommended for DNA extraction from stool, and almost allof them are difficult and time consuming, dealing with high amount of toxic materials like phen...

متن کامل

DNA Extraction of Almond without Phenol and Liquid Nitrogen

Genomic DNA extraction with a high quantity and quality is a basic requirement in molecular biology. The DNA obtained was free of any contamination proteins, polysaccharide, polyphenols and colored pigments. These compounds would interfere with the genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The isolated genomic DNA was fo...

متن کامل

An efficient and simple CTAB based method for total genomic DNA isolation from low amounts of aquatic plants leaves with a high level of secondary metabolites

An efficient DNA isolation protocol specifically modified to get pure quality DNA required for molecular studieshas been reported in this paper. Some aquatic plants (Potamogeton spp., Ceratophyllum demersum and Myriophyllum spicatum) were used for the study. The protocol developed will be useful in getting high and pure DNA. Instead of using the available DNA extraction kits, this protocol can ...

متن کامل

Optimization of the genomic DNA extraction in some mosses

The presence of organic compounds and high amount of secondary metabolites (polysaccharides, phenolic component, etc.) in mosses cause difficulties in DNA extraction that are followed by problems in PCR reactions. In lower plants, various methods have been used for DNA extraction including silica gel and different commercial kits. These methods mostly use hazardous (like phenol or liquid nitrog...

متن کامل

An optimized affordable DNA-extraction method from Salmonella enterica Enteritidis for PCR experiments

In diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. If not inactivated properly, cellular and/or environmental DNA nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in PCR experiments. When crude cell extracts, proteinase K–treat...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Genetics and molecular research : GMR

دوره 10 1  شماره 

صفحات  -

تاریخ انتشار 2011