Epicentre
MasterPure™ RNA Purification Kit
MCR 85102
Isolation and handling of ribonucleic acid (RNA) is more difficult than DNA due to the presence of ubiquitous RNases. Purification of high-quality, intact RNA from biological samples or in vitro reactions is important to the success of all downstream applications. To achieve reproducible, trustworthy results from quantitative RT-PCR, Northern blots, microarrays, and cDNA library prep for Next-Generation sequencing, it is important to use the best RNA purification methods available.
To check the expression pattern of genes of interest in real time-PCR.
SD rats' liver tissue
I have used the kit to isolate RNA from liver tissue. We then collected the liver tissue after immediate scarified animals. Next, we homogenized fresh tissue and added to micro centrifuge tube. Added 300 µl of tissue lysate to that, added Proteinase K and mixed well. This was incubated at 65°C for 20 minutes. From there, we followed the manufactures’ instructions.
Take care that the tissues are always ground with liquid nitrogen only.
RNA A260/A280 ratio in between of 1.7-2.0 with anodrop
None
With less contamination got good yield of RNA.
Columns are expensive.
Protocol is simple and yields good results.