The E.Z.N.A.® HP Total RNA Isolation Kit provides a rapid and easy method for RNA isolation from a small amount of cultured eukaryotic cells or tissues. This kit allows single or simultaneous processing of multiple samples in less than 40 minutes. Normally, 1 x 107 eukaryotic cells or 25-30 mg tissue can be used in a single experiment. There is no need for phenol/chloroform extractions, time-consuming steps such as CsCl gradient ultracentrifugation, or precipitation with isopropanol or LiCl. RNA purified using the E.Z.N.A.® HP Total RNA method is ready for applications such as RT-PCR, RT-qPCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
For Research Use Only. Not for use in diagnostic procedures.
|Downstream Application||PCR, qPCR, real-time RT-PCR, microarray, Northern blot, poly-A purification, RNA-Seq|
|Elution Volume||40-70 μL|
|Starting Material||Cultured eukaryotic cells and soft tissues|
|Starting Amount||1 x 107 eukaryotic cells or 25-30 mg tissue|
|RNA Yield||Up to 100 µg|
|Processing Mode||Manual, centrifugation/vacuum|
|Throughput||1 - 24|
|RNA Binding Technology||Silica Mini spin column|
|Binding Capacity||100 µg|
Protocol and Resources
Product Documentation & Literature
The yield of total RNA purified from tissues and cells using E.Z.N.A.® HP Total RNA Kit was higher than using a competing product.
Figure 1. RNA yields obtained using the Omega and Qiagen kits with tissue samples and cultured cell samples. RNA yields were 18.36 μg and 28.17 μg from the mouse kidney, 17.32 μg and 19.96 μg from the HEK293 cells using Qiagen and Omega Bio-tek kits respectively. One-way ANOVA followed by Tukey’s post-hoc analysis indicates that the RNA yield extracted using Omega Bio-tek’s kit significantly better than that of Qiagen’s (p < 0.01) for both the sample types tested.
RNA Integrity Analysis – Tissue Samples
Figure 2. Purified RNA from tissue samples were analyzed on an Agilent Tapestation 2200 to provide integrity along with gel images. a) Representative electropherogram – Qiagen; b) representative electropherogram – Omega Bio-tek.
RNA Integrity Analysis – Cultured Cells
Figure 3. Purified RNA from cultured cells were analyzed on an Agilent Tapestation 2200 to provide integrity along with gel images. a) Representative electropherogram – Qiagen; b) representative electropherogram – Omega Bio-tek.
gDNA Contamination of Isolated RNA
Figure 4. Genomic DNA contamination in the isolated DNA. With the RNeasy Plus Kit, DNA co-eluted was found to be 4.52% of total RNA for both the sample types tested. And it was 4.79% for HP Total RNA Kit for the same. The findings suggest that the RNA isolated from both the kits was of high quality with appreciable gDNA removal.
RNA Yield and Quality from Cultured Cells
Figure 5. RNA was purified from 1 x 106 cultured HEK293 Cells with the E.Z.N.A. HP Total RNA Kit. RNA concentration determined by optical density measurements with NanoDrop® 2000c. Total elution volume was 100 µL.
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