Archiving of formalin-fixed, paraffin-embedded (FFPE) tissue sections represents a valuable and widely sourced biomedical research material. As more and more researchers turn to molecular analysis of FFPE samples to develop specific operational steps, it is becoming increasingly important to consider the unique properties of these samples. QIAGEN offers a complete solution for the entire process of FFPE sample purification and downstream testing. QIAamp DNA FFPE Tissue Kit - Purification of genomic DNA from paraffin-embedded tissues Specially optimized lysis and reverse cross-linking Buffer to ensure efficient release of DNA Quickly purify high quality, ready-to-use DNA from wax blocks in 30 minutes Flexible elution volume: from 20 μl to 100 μl A single sample size can be as high as 8 wax block samples Automated purification on QIAcube Genomic DNA was purified from different FFPE tissue samples using the QIAamp DNA FFPE Tissue Kit. Real-time quantitative PCR amplification of prnp gene (78 bp, 464 bp and 727 bp) products Figure 1 PCR analysis of DNA purified from FFPE tissue samples. RNeasy FFPE Kit - Purification of total RNA from paraffin-embedded tissues Optimized cleavage and reverse cross-linking buffers ensure efficient release of RNA Fast operation, purified RNA in 85 minutes Contains DNase to ensure contamination without genomic DNA Automated purification on QIAcube After removing the paraffin from the sample, the enzyme cleavage was performed in the optimized lysis buffer, and the sample cleavage was completed in 15 minutes without affecting the RNA yield. After lysis, the samples were incubated at 80 °C for 15 minutes to reverse formalin cross-linking. Genomic DNA was then efficiently removed using DNase and DNase Booster Buffer, including small DNA fragments in FFPE samples. Finally, the concentrated RNA was purified using an RNeasy MinElute spin column. The elution volume of RNA is only 14–30 μl, thus reducing the volume of reaction in downstream operations. Figure 2 RNeasy FFPE experimental procedure. Total RNA was purified from rat kidneys using the RNeasy FFPE Kit. Real-time RT-PCR analysis of PGK1 (phosphoglycerate kinase 1) was performed on a Rotor-Gene Q real-time PCR analyzer using (+RT) or without (-RT) reverse transcriptase. The -RT curve demonstrates that RNA purified using the RNeasy FFPE Kit contains virtually no genomic DNA. miRNeasy FFPE Kit - Purification of total RNA containing miRNA from paraffin-embedded tissues Effective purification of miRNA and total RNA (see Figure 3) Optimized cleavage and reverse cross-linking buffers ensure efficient release of RNA Contains DNase to ensure contamination without genomic DNA Figure 3 Successful real-time RT-PCR analysis from FFPE samples. Rat liver tissue was fixed with formalin for 24 hours or 60 hours, and then total RNA including miRNA was purified using the miRNeasy FFPE Kit. Quantitative real-time RT-PCR analysis was performed using the miScript PCR System and purified RNA as a template to detect larger mRNAs of miRNA miR-16 and miR-29a and PGK1 genes. The results showed that both miRNAs and mRNAs were detected simultaneously from the same eluate. Product List: product name Item number QIAamp DNA FFPE Tissue Kit (50) 56404 RNeasy FFPE Mini Kit (50) 74404 miRNeasy FFPE Kit (50) 217504 QIAamp DSP DNA FFPE Tissue Kit (50) 60404 Deparaffinization Solution (16 ml) 19093 Figure 4 Efficient real-time RT-PCR. Super Absorption Natural Diatomite Non Slip Bath Mat .Its main material is diatomaceous earth.Its main characteristic is super super super absorbent,which can instantly absorb water from your feet and non-slip.It is also all-natural and can absorb harmful chemicals from the air 24 hours a day.What's more, it's easy to clean up and can be customized. easy life Anti Slip Floor Mat,Printing Bath Mat Carpet,Diatomaceous Mat Homeware,Fast Drying Carpet Diatomite zhangjiagang wellyoung material co.,ltd , https://www.wellyoung-board.com
August 10, 2021