P0907 Organic Extraction of RNA From Plant and Animal Tissues Without Phase Separation

Stanislav Forman , Zymo Research Corp., Irvine, CA
Danice Cabaya , Zymo Research Corp., Irvine, CA, Irvine, CA
Xi Yu Jia , Zymo Research Corp., Irvine, CA
Organic acid-guanidinium-phenol based extraction is widely implemented for most plant and animal RNA purification protocols.  The organic extraction method effectively inactivates nucleases and infectious agents and allows for adequate sample storage and stabilization. However, limitations of existing organic extraction methods become apparent with high-throughput processing and the handling of small volume inputs.  In both cases, the requirement of phase separation can affect greatly both the yield and purity of RNA.

Here we present an alternative acid-guanidinium-phenol based procedure that effectively bypasses phase separation/precipitation steps with a spin column and specially designed washes.  This helps to eliminate problems attributed to phenol carryover that is often associated with conventional organic extraction methods.  This new “Direct-zol” procedure maximizes total RNA recovery, including small RNAs, without the need for a carrier.  For tough-to-lyse plant and animal tissue the Direct-zol procedure can be combined with our unique BashingBead™ and OneStep inhibitor removal technologies for unparalleled yields and complete removal of polyphenolic inhibitors to RT-PCR, respectively.  The Direct-zol procedure delivers DNA-free RNA that is ideal for RT-PCR, RNA-seq (expression profiling), hybridization, etc.