RecoverAll™ FFPE Total Nucleic Acid Isolation Kit for isolation of formalin or polyformalin-fixed, paraffin-packed nucleic acids Total nucleic acids were extracted from buried (FFPE) tissues. Includes sufficient reagents for 40 purifications each from up to four 20 µm sections or up to 35 mg of unsectioned core sample. Features of the RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE:

• Optimized for isolation of total nucleic acids, including microRNA, from FFPE tissue
• No overnight proteinase K digestion required - dewax in the morning, qRT-PCR in the afternoon
• Typical yield> 50% of unfixed tissue
• Recovered nucleic acids are suitable for next generation sequencing, real-time RT-PCR, PCR, mutation screening and microarray analysis
< br>Extracting nucleic acids from disparate samples
Archived tissue samples contain valuable disease state information, but it has traditionally been difficult to separate nucleic acids from samples of a quality suitable for molecular analysis. Standard preservation techniques use formalin, which preserves tissue structure and prevents perishability, but also captures nucleic acids and modifies them through protein-protein and protein-nucleic acid cross-linking. RNA (and to some extent DNA) is often fragmented and chemically modified in this way, making it incompatible with many molecular analysis techniques. RNA fragmentation in FFPE tissue cannot be reversed; however, the protease digestion conditions of the RecoverAll™ Kit are designed to release large amounts (as shown) of all sizes of captured RNA fragments, including microRNA, in a relatively short time.

RecoverAll™ Total Nucleic Acid Isolation Kit Solution
RecoverAll™ Total Nucleic Acid Isolation Kit operation requires approximately 45 minutes of hands-on time and can typically be completed in less than 1 day Complete the operation on RNA within a certain time. FFPE samples were dewaxed using a series of xylene and ethanol washes. Next, they undergo extensive protease digestion, and the incubation time should be consistent with the requirements for RNA or DNA recovery. The nucleic acids are then purified using fast glass filtration and eluted into water or low-salt buffer.

Nucleic acids for virtually any downstream application
As is true for all FFPE tissues, sample fixation and storage often results in nucleic acid fragmentation and modification. Therefore, downstream applications (such as microarray analysis, which require more raw RNA than qRT-PCR) may require modifications to obtain optimal results. Although DNA generally does not fragment as easily as RNA, it is more reactive with formalin and requires a longer (2 days) protease digestion time to release large amounts of DNA.