Hiiii
Can anyone advice regarding the best kit for RNA isolation from urine samples?
I have used qiagen viral RNA kit but it gives vey high O.D ( ~ 3.1 )
I need purified RNA with good quality so that it can be used for real time PCR.
What would be better option to isolate RNA either from supernatant or cell pellet ??
Hi,
I recommand the ZR Urine RNA Isolation Kit (Zymo research). Best Luck.
Total RNA is isolated with the use of a commercial kit (RNeasy mini kit, Qiagen, Valencia, California). In brief, 350 Al of RLT buffer (Qiagen) is added to the urine cell pellet and 450 Al of the buffer is added to the biopsy specimen. RNAlater (50 Al) is added to the renal biopsy samples just prior to RNA isolation; 50 Al of RNAlater is also added to the urine cell pellets that are not treated with RNAlater prior to snap freezing. The urine cell pellets/renal tissues are homogenized using a 1-ml syringe and a 23-gauge needle (cell pellet) or rotor homogenizer (biopsy tissue). The homogenate is passed through a QIA Shredder column (Qiagen) to remove insoluble debris. RNA amount is measured with the use of a UV spectrophotometer and absorbance is measured at A260 and A280 nm and expressed as A260/A280 ratio.
see this link
http://www.gmo-qpcr-analysis.info/medeiros-rna-2003.pdf
- Badges
- Science topic