EXTRACTME TOTAL RNA MICRO SPIN KIT (EM31.1)

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.

PRODUCT SPECIFICATIONS
• SAMPLE MATERIAL
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
BINDING CAPACITY
Approx. 30 μg RNA
• TIME REQUIRED
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
• RNA PURITY
A260/A280 ratio = 1.7 – 1.9

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