EXTRACTME TOTAL RNA KIT, 250 preps supplied by BLIRT is intended to ensure a quick and effective purification of high quality RNA from up to 30 mg of tissue (fresh or frozen) and up to 107 cultured cells. The protocol of isolation and buffer formulations was optimized to ensure high efficiency of isolation and purity of RNA. 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 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, RT-qPCR, Northern blotting, cDNA synthesis, primer extension, RNA sequencing, microarrays.

Product Specifications:

Product name - EXTRACTME TOTAL RNA KIT, 250 preps

Catalog No - EM09.2-250

Brand: BLIRT

Pack size: 250 preps

Product Features:

1SAMPLE MATERIAL

fresh or frozen tissue : up to 30 mg

tissue preserved in RNase inactivating buffers (e.g. RNAlater, Ambion): up to 30 mg

cell culture: up to 107 cells.

2. BINDING CAPACITY

~ 230 g RNA

3. DNA FRAGMENT LENGTH

100 bp 10 kb

DNA fragments in the range of 50100 bp and 1020 kb can also be purified as can genomic and plasmid DNA, however the efficiency will be decreased.

4. TIME REQUIRED

1012 minutes (lysis and homogenization time not included)

1530 minutes for homogenization in liquid nitrogen

1520 minutes for mechanical homogenization (ceramic beads)

5 minutes for optional DNase I treatment

5. RNA PURITY

A260/A280 ratio = 1.9 2.1