For BiOstic® FFPE Tissue DNA Isolation Kit click here
The BiOstic® FFPE Tissue RNA Isolation Kit is optimized for complete lysis and extraction of total RNA from formalin-fixed, paraffin-embedded tissues without the use of organic solvents. A proprietary buffer formulation in conjuction with a Proteinase K lysis facilitates complete dissolution of the wax during a 60°C heating step to release the tissue. A second 70°C heating step removes the cross-links that are inhibitory to RT-QPCR. RNA from the melted sample is purified on our new low elution silica spin filters to increase the final RNA concentration. Genomic DNA is removed in an on-column DNase digestion step utilizing our room temperature stable (RTS) DNase. RNA from the BiOstic® FFPE Tissue RNA Isolation Kit it ready to use in any downstream application including RT-QPCR, microarrays and sequencing.
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Fig 1. The fixation process and storage of FFPE samples frequently leads to nucleic acid degradation and base modification. DNA prepared with the BiOstic® FFPE Tissue DNA Isolation Kit (MB) shows higher molecular weight distribution when compared to the competitor’s xylene protocol (Q).
Fig 2. Comparative analysis of the BiOstic® FFPE Tissue RNA Isolation Kit (MB) and competitors Q and LT of RNA extraction from a single 10 micron tissue slice of FFPE Normal Human Liver tissue. a) 1.2% TAE gel showing higher yields of intact RNA obtained when sample was prepared with the BiOstic® FFPE Tissue RNA Isolation Kit. b) Invitrogen Qubit™ Fluorometer readings show that higher yields of RNA were obtained when sample was prepared with the BiOstic® FFPE Tissue RNA Isolation Kit (MB).
Fig 3. Successful RT-qPCR amplification. (a) RNA described in Fig 2 was analyzed by RT-qPCR using a TaqMan® ActB assay. Samples prepared with the BiOstic® FFPE Tissue RNA Isolation Kit were free of inhibitors, as indicated by successful amplification and a difference of approximately 3 cycles between the undiluted RNA (Dark Blue) and the 1:10 dilution (Light Blue). (b) Undiluted samples prepared with the Qiagen RNeasy FFPE Kit (Green) and Life Technologies PureLink® FFPE Total RNA Isolation Kit (Brown) demostrated inhibition.
|Format||Silica Spin Filter Tubes|
|Method||Direct melting of FFPE tissues|
|Sample Size||1 to 5 single slices or 15 mg of tissue|
|Binding Capacity||Up to 6 µg per spin filter|
|RNA Isolated||Total RNA|
|Equipment Required||Vortex, centrifuge, and water bath or heat block|
Store the Low Elution Spin Filters at 4°C.
Proteinase K (Solution FR2) and the RTS DNase are stable at room temperature for up to 6 months. For prolonged storage (up to 2 years), place the Solution FR2 and RTS DNase at 4°C.
Store all other reagents and kit components at room temperature (15-30°C).