EXTRACTME miRNA KIT, 50 preps supplied by BLIRT is intended to ensure a quick and effective purification of miRNA with chances of purification of large RNA and DNA simultaneously. The protocol of isolation and buffer formulations was optimized to ensure high efficiency of isolation and purity of miRNA. The process for RNA purification uses spin minicolumns with membranes which bind nucleic acids efficiently and selectively at high concentration of chaotropic salts. The 1st isolation step involves the homogenization of a tissue which results in the disintegration of intercellular bonds (epithelial tissue) and fragmentation of high-molecular proteins (muscle or connective tissue). Lysing of the homogenate is done with guanidine thiocyanate and detergents. Guanidine thiocyanate and -mercaptoethanol (optional) is utilized to inactivate RNases. The separation of the homogenate from undigested tissue/cell that remains after centrifugation is done. The RNA then binds to the Purification Column membrane by selective conditions in mixture after adding ethanol. On-column DNase digestion enables removal of remaining genomic DNA. A three-step washing stage adequately eliminates contaminations and protein inhibitors. By the help of low ionic strength buffer or RNase-free water the purified RNA is eluted out that may be utilized in several downstream applications, like RT-PCR, Northern blotting, RT-qPCR and so forth.

Product Specifications:

Product name - EXTRACTME miRNA KIT, 50 preps

Catalog No - EM12-050

Brand: BLIRT

Pack size: 50 preps

Product Features:

1 SAMPLE MATERIAL

fresh or frozen tissue (stored at -80C): 1 10 mg

tissue preserved in RNase inactivating buffers: 1 10 mg

cell culture: 104 104 cells

2. BINDING CAPACITY

~ 90 g

3. miRNA TIME REQUIRED

25 30 minutes (lysis and homogenization time not included)

35 70 minutes for homogenization in liquid nitrogen

35 50 minutes for mechanical homogenization (ceramic beads)

4. RNA PURITY

A260/A280 ratio = 1.9 2.1