I know that TRIzol or QIazol can be used in combination with chloroform and phase lock gels to isolate good quality RNA from impurities and DNA, but apparently RNAzol RT which, has been used for q RT-PCR studies does not require the chloroform and phase separation steps. Anybody care to clarify this? Thanks.
I use RNAzol for all of my RNA extractions and it works great. I have done numerous RNA-Seq and qRT-PCR studies with the extracted RNA with no problems. The RNA is very pure in terms of 260/230 and 260/280 and usually have RIN values around 9. To make things even easier, you can combine the RNAzol with the Direct-zol kit from Zymo and then no phase separation is required. Good luck.
Zach
Using RNeasy mini kit (Qiagen) with DNase treatment can results in faster but purified high quality RNA.
I am working with buffalo/goat mammary tissues that I bring from slaughterhouse in
RNAlater and 4 deg C overnight. Next day I process it- take one piece (~50 mg) and put into RNAzol and store at -80 deg C. I proceed with homogenization of sample with hand held tissue homogenizer for 1-2 min. After addition of 0.4 ml of nf water/ml of RNAzol, I centrifuge samples to obtain supernatant. Surprisingly, I do not see any RNA pellet in supernatant after addition and incubation with equal volume of isopropanol. Running 1% gel does not make me happy, however, OD ratios often look OK. When I replace isopropanol with 75% chilled ethanol (made in nf water) I do see pellet (of mRNA and miRNA) better. Why it is happening? Where do I am going wrong?
Any help in this will be helpful.
hi we have problem with RNA extration from whole blood with RNAzol from MRC can someone help me what can be wrong?
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