35,00 € – 691,00 €
RNA isolation - EXTRACTME TOTAL RNA MICRO SPIN KIT is designed for the rapid and efficient purification of high quality RNA up to 15 mg of tissue (fresh or frozen) and up to 106 cultured cells with an extremely low elution volume of only 5 μl. The isolation protocol and buffer formulations were optimized for high isolation efficiency and purity of RNA. The product is intended for research use only.
The EXTRACTME TOTAL RNA MICRO SPIN KIT utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids at high concentration of chaotropic salts. In the first isolation step, the tissue is homogenized in order to disintegrate intercellular bonds (epithelial tissue) and fragmentize high-molecular proteins (muscle or connective tissue). Then the homogenate is lysed with guanidine thiocyanate and detergents. Any RNases are inactivated by guanidine thiocyanate. The homogenate is separated from the undigested tissue/cell remains by centrifugation and on the Homogenizing Column. The RNA is bound to the Purification Micro Spin Column membrane by addition of ethanol. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified RNA is eluted using a low ionic strength buffer or RNasefree water (pH 7.0-9.0) and can be used directly in all downstream applications such as RT-PCR, Northern blotting, RT-qPCR and so forth.
- fresh or frozen tissue (stored at -80°C): up to 15 mg
- tissue preserved in RNase inactivating buffers: up to 15 mg
- cell culture: up to 106 cells
Approx. 30 μg RNA
- 16-20 minutes (lysis and homogenization time not included)
- 30-60 minutes for homogenization in liquid nitrogen
- 30-40 minutes for mechanical homogenization (ceramic beads)
- 15 minutes for optional DNase I treatment
A260/A280 ratio = 1.7 – 1.9
RECOMMENDATIONS AND IMPORTANT NOTES
EXTRACTME TOTAL RNA MICRO SPIN KIT (EM31) - Manual
EXTRACTME TOTAL RNA MICRO SPIN KIT – TROUBLESHOOTING
The quality of each production batch (LOT) of the EXTRACTME TOTAL RNA MICRO SPIN KIT is tested using standard QC procedures. The purified RNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by reverse transcription and qPCR.