Total RNA Kit, peqGOLD

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- This kit purifies RNA from up to 100ug tissue	- All steps must be carried out at room temperature (22 – 25 °C). - Pre-heat RNase-free Water to 70 °C prior to elution. - For DNA contamination Digest with Rnase free DNase and incubate at 37 °C for 5 min.	- Ensure RNA Wash Buffer II Concentrate has been diluted with 4 volumes of 100 % ethanol as indicated on the bottle for ant salt carryover during elution

Upstream tips
- This kit purifies RNA from up to 100ug tissue
Protocol tips
- All steps must be carried out at room temperature (22 – 25 °C). - Pre-heat RNase-free Water to 70 °C prior to elution. - For DNA contamination Digest with Rnase free DNase and incubate at 37 °C for 5 min.
Downstream tips
- Ensure RNA Wash Buffer II Concentrate has been diluted with 4 volumes of 100 % ethanol as indicated on the bottle for ant salt carryover during elution

Publication protocol

Total RNA was extracted from HGT-1 cells and two human biopsy specimens by using the peqGold Total RNA Kit (Peqlab). Quantity and quality were checked spectrophotometrically. Reverse transcription was carried out with 2 µg RNA and the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). Real-time PCR was performed with an Applied Biosystems StepOneplus Real Time PCR system and Fast SYBR Green Master Mix (Thermo Fisher Scientific). Primers were designed using the National Center for Biotechnology Information (NCBI) primer designing tool (using Primer 3 and BLAST) . Cycling conditions were 20 s/95 °C (activation), 3 s/95 °C (denaturation), 30 s/60 °C (annealing), and 15 s/67 °C (elongation with fluorescence measurement)

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Manufacturer protocol

Download the product protocol from VWR for Total RNA Kit, peqGOLD below.

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