GeneAll® Exgene™ FFPE Tissue DNA kit provides a convenient and easy method for the isola-tion of total DNA from Formalin Fixed and Paraffin Embedded (FFPE) specimen by non-organic solvent. FFPE is one of the most commonly used methods of clinical tissue preservation; the clini-cal tissue is fixed by formalin and subsequently embedded in paraffin to keep its original form.
The FFPE tissue is useful in disease research such as microscopic observation and immunohisto-chemical analysis. And the extracted-nucleic acid from FFPE specimen can be used for molecular diagnosis of various diseases. However, during the fixative process, the nucleic acids in FFPE are damaged significantly by various degrees of crosslinking between DNA and protein, and the dam-age get worse during its long-term preservation. For such a reason, the DNA isolated from the preserved FFPE specimen generally has low qualities in its yield, purity, integrity and PCR-proces-sivity. But despite these problems, the purified nucleic acids from FFPE specimen are widely used for the PCR targeted to relatively short DNA fragments.
To obtain DNA from FFPE tissue by GeneAll® Exgene™ FFPE Tissue DNA kit, FFPE specimen is deparaffinized in buffer DP which rapidly separate tissue from paraffin sections, and then the sam-ple is lysed in the optimized buffer containing detergents and lytic enzymes. Under high salt condi-tion, DNA in the lysate binds to silica membrane and impurities pass through membrane in to a collection tube. The membrane is washed with a series of alcohol-containing buffer to remove any traces of proteins, cellular debris and salts. Finally pure DNA is released into a clean collection tube with deionized water or low ionic strength buffer.
This fast and convenient procedure of GeneAll® Exgene™ FFPE Tissue DNA kit takes only 150 min for complete preparations of total DNA. Purified DNA can be used directly for PCR (≤500 bp), real-time PCR, and other downstream applications.