Aurum™ Total RNA Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- This kit is designed to process upto 2x10^6 cells. - Before using lysis solution, add 500 ul of β-mercaptoethanol (β-ME) to the solution for a final concentration of 1%.	- To perform efficient elution preheat the elution solution at 70C in water bath priorto the elution step. - If there is high genomic DNA contamination, increase DNAse I digestion time and use only the DNAse dilution solution provided in the kit.	- For better downstream application add appropriate volume of 95-100% ethanol to the wash solutions before intial use. - If elute volume is >80ul, there is a possibility of ethanol contamination. To reduce this, add 1-3min of centrifugation time after the final wash step.

Upstream tips
- This kit is designed to process upto 2x10^6 cells. - Before using lysis solution, add 500 ul of β-mercaptoethanol (β-ME) to the solution for a final concentration of 1%.
Protocol tips
- To perform efficient elution preheat the elution solution at 70C in water bath priorto the elution step. - If there is high genomic DNA contamination, increase DNAse I digestion time and use only the DNAse dilution solution provided in the kit.
Downstream tips
- For better downstream application add appropriate volume of 95-100% ethanol to the wash solutions before intial use. - If elute volume is >80ul, there is a possibility of ethanol contamination. To reduce this, add 1-3min of centrifugation time after the final wash step.

Publication protocol

After respective treatments, total RNA from BMVEC monolayers was harvested and prepared by using the Aurum Total RNA Extraction Kit (Bio-Rad, CA). RNA was reverse-transcribed using the iScript cDNA Synthesis Kit (Bio-Rad, CA), using 1 µg of total RNA per 20-µl reaction. Validated primers (PrimePCR Assays; Bio-Rad, CA) for the genes and reference genes that were quantified in these studies were used in either individual assays or as custom-designed PCR Arrays (PrimePCR Arrays; Bio-Rad, CA). G6pdx and Actb were used as reference genes. For real-time PCR reactions, SsoAdvanced SYBR Green Supermix was used, and RT-PCR was run and analyzed on Bio-Rad CFX Connect (Bio-Rad, CA) as per the manufacturer's instructions.

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Manufacturer protocol

Download the product protocol from Bio-Rad Laboratories for Aurum™ Total RNA Mini Kit below.

Download manufacturer protocol

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