Total RNA
Mini Kit
Total RNA Mini Kit is designed for a rapid and efficient purification of high quality RNA from animal tissue, bacterial cells, yeast, fungi or viruses. The isolation protocols and buffer formulations were optimized for high isolation efficiency and purity of RNA.
Description
RNA purification procedure utilizes spin mini columns with membranes which efficiently and selectively bind nucleic acids at high concentration of chaotropic salts. During the first isolation step, a tissue is homogenized in order to disintegrate intercellular bonds (epithelial tissue) and fragmentize high-molecular proteins (muscle or connective tissue). A homogenate is lysed with guanidine thiocyanate and detergents. RNases are inactivated by guanidine thiocyanate and ß-mercaptoethanol (optional). The homogenate is separated from undigested tissue/cell that remains after centrifugation. RNA binds to a mini column membrane by addition of ethanol. A three-step washing stage effectively removes impurities and enzyme inhibitors. Purified RNA is eluted with the use of low ionic strength buffer or RNase free water and may be used directly in all downstream applications, such as RT-PCR, RT-qPCR and cDNA synthesis.
REF. NUMBER
155NAIS0401
SAMPLE MATERIAL
> fresh or frozen solid tissue
> animal and bacterial cell culture
> yeast and fungi
> plasma, serum, cell-free body fluids, and culture supernatants
PROTOCOL EXECUTION TIME
25-35 minutes (including incubation time)
ADDITIONAL REAGENTS AND EQUIPMENT REQUIRED
> 96–100% ethanol PFA
> 1.5–2.0 ml sterile microcentrifuge tubes
> microcentrifuge with rotor for 1.5–2.0 ml (≥ 11,000 x g)
> vortex mixer
> racks for 1.5 and 2.0 ml microcentrifuge tubes
> automatic pipettes and pipette tips (in range 2-20, 20-100, and 100-1000 μl)
> disposable gloves
Might be necessary:
> tweezers, scalpel
> mechanical homogenizer
> 100% ß-mercaptoethanol
> Lyticase
> 1M DTT
QUANTITY
50 isolations
CERTIFICATES