STAR BEADS RNA Extraction kit
STAR BEADS RNA Extraction kit
Rapid isolation of total RNA from various biological samples
Overview
STAR BEADS RNA Extraction Kit provides a fast and efficient purification method to isolate high-quality total RNA from as few as 15 cells up to 2 x 106 cells.
The isolated RNA is highly pure and ready for downstream reactions, including real-time RT-PCR, RNA-seq, and other enzymatic reactions.
STAR BEADS RNA Extraction Kit is available in bottle format for 96 tests.
The kit is fully automatable in a variety of extraction instruments. For automated scripts and protocols, visit the document section on our website www.cyanagen.com or email to technical.support@cyanagen.com
Typical output of RNA extraction using STAR BEADS RNA Extraction Kit
Total RNA from 106 ovarian cell line was isolated with STAR BEADS RNA Extraction Kit and separated by gel electrophoresis on a 1% agarose gel.
Features
• Co-purification of short and long RNA: messenger RNA, ribosomal RNA, miRNA and other small RNAs are isolated in the same extraction
• No genomic DNA contamination
• Quantitative RNA recovery from 15 – 2,000,000 cells
• Ultra pure RNA: absence of PCR contaminants in the eluates, ready for downstream applications
• Eco-friendly: Magnetic bead-based technology allows up to 50% reduction in plastic use
• Rapid: 85 minutes protocol time for manual and automatic extraction
• Easy storage: room temperature
• Automatable: compatible with
– Allsheng Auto-Pure96/Auto-Pure32A/Mini
– Accuris IsoPure™ Mini/96
– Procomcure Phoenix-Pure96/32
– MOLGEN PurePrep 96/32
– BIOER GenePure Pro NPA-32P
– MGI SP-NE32
– BIGFISH BFEX-32
…and many other systems
Kit Specifications and typical output
Qunatitative results on RNA from cells over a 5 Log range.
RNA was extracted from 2 millions HCT116 cells down to 15 cells. rRNA 18S gene was amplified by realtime RT-qPCR on the RNA samples. Quantitation standard curve is obtained plotting threshold cycle values (Cq) against the log of cell number in the input. Optimal correlation is demonstrated by R2 = 0.99.
Maximal recovery across a range of elution volumes.
RNA yield is consistent between 30µl and 100µl of elution buffer volumes. Purity (A260/280) is unaffected by elution volume. Data represent mean ±SD, N=2. Standard elution volume is 100 µl, however RNA can be eluted in down to 30 µl for a more concentrated solution.
Linear recovery of RNA from 105 to 2 x 106 cells.
RNA was extracted from 105, 2 x 105, 5 x 105, 106 and 2 x 106 HCT116 cells. The concentration of the extracted samples were measured by a nanophotometer (IMPLEN N80) and reported versus the sample amount, affording a linear correlation with a R2= 0.97.
Amplification of both large and small RNAs from the same sample.
Total RNA was isolated from 2,5 x 105 cells and RT-qPCR was performed on a human microRNA (miR-16), a small nucleolar RNA (U6), a large mRNA (GAPDH) and a human rRNA (18S). Data represent results (mean ± SD, N=4) using STAR BEADS RNA Extraction Kit and various silica-based (Competitor Q) and magnetic beads-based competitors (Competitor MN and Competitor TS) following manufacturers’ instructions. Statistical significance is indicated with * when p < 0.05, with ** when p < 0.01, with *** when p < 0.001.
Excellent RNA Integrity and isolation of large and small species of RNAs.
RNA was isolated using Cyanagen STAR BEADS RNA Extraction Kit (A), Competitor MN (B), Competitor TS (C), Competitor Q (D). One microliter of the 100 µL isolated RNA was analyzed on the Agilent® 2100 BioAnalyzer using Agilent RNA 6000 Nano kit chip. The integrity of RNA extracted with Cyanagen STAR BEADS RNA Extraction Kit shows the presence of small RNA species. Cyanagen STAR BEADS RNA Extraction Kit consistently isolates high quality RNA with RNA Integrity Number (RIN) always > 8.5.
Excellent RNA Integrity.
Total RNA was extracted from 2.5 x 105 human ovarian cancer cells OC314 (lane 1-2) and from 2.5 x 105 human colon cancer cells HCT116 (lane 3-4) using the STAR BEADS RNA Extraction Kit and analyzed using Bioanalyzer RNA Analysis – Agilent RNA 6000 Nano Kit on Agilent 2100 Bioanalyzer™.
Panel A. Gel-like image generated by Bioanalyzer™ showed excellent RNA integrity with RNA Integrity Number (RIN) always > 8.5 for all the cell samples analysed.
Panel B. Electropherogram of sample #3 showed two very clear band corresponding to 18S and 28S. rRNA Ratio [28S / 18S] is 3.2 and RNA Integrity Number (RIN) is 9.9 .
Absence of PCR inhibitors in eluates.
RT- qPCR of GAPDH mRNA amplified in 6 ten-fold dilutions of RNA samples extracted from 2.5 x 105 HCT cells STAR BEADS RNA Extraction Kit. Quantitation standard curve is obtained plotting threshold cycle values (Ct) against the log of template RNA concentration. An optimal PCR efficiency is observed (E= 100,1%, slope= -3,320, R2 = 0,997) indicating the absence of PCR inhibitors.
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