SV 96 Total RNA Isolation System
The SV 96 Total RNA Isolation System provides a fast and simple method for the preparation of intact, purified total RNA from tissue culture cells. The system provides a
fast and simple technique for the preparation of purified, intact RNA that allows automation on liquid handling workstations such as the Beckman Biomek® 2000 or FX. It is a complete “walk-away” method that can be completed in less than one hour per plate. The automated isolation method reproducibly purifies high-quality RNA that may be amplified by RT-PCR. The flexible format of SV 96 Total RNA Isolation System can also be performed manually.
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Figure 1. Total RNA isolation and purification using the SV 96 Total
RNA Isolation System.
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The SV 96 Total RNA Isolation System incorporates an on-membrane DNase treatment step to substantially reduce genomic DNA and allows recovery of total RNA from tissue culture cells. The unique design of the system eliminates waste handling and cross-contamination between samples.
The procedure of SV 96 Total RNA Isolation System purifies total RNA from cell lysates using 96-well vacuum filtration steps, which eliminates the need for centrifugation. Isolation of RNA can be used from different materials including; high-throughput from tissue culture cells, mouse liver lysate and plant tissue
lysate. |
FEATURES
Convenience
Ready-to-use plates and solutions provide convenience and speed. RNA from samples in
96-well plate can be purified in less than one hour.
Choice of Automated or Manual Formats
Purify total RNA manually or automate for walk-away operation.
Ready for RT-PCR
RNA purified with the system is ready for use in RT-PCR without further processing.
The SV 96 Total RNA Isolation System provides total RNA ready for direct use in RT-PCR. In this example, RNA was isolated from known amounts of SH-SY5Y cells and eluted in ~70µl of Nuclease-Free Water. Four microliters of eluate was then used in the Access RT-PCR System
(Cat. # A1250) with ß-Actin Primers
(Cat. # G5740).
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Figure 2. Limit of detection of total RNA purified from 2X dilution series of human neuroblastoma cells. Total RNA was isolated from a human neuroblastoma cell line (SH-SY5Y cells) using the SV 96 Total RNA Isolation System. Four microliters of eluted RNA was amplified using ß-actin primer pairs. PCR products were run on a 2% agarose gel and visualized by ethidium bromide staining. Lane headings indicate the number of cells per isolation, from 50,000 to 24. Signal is clearly present down to 391 cells. |
Consistent Yield - Yield is consistent across the entire plate.
The SV 96 Total RNA Isolation System provides consistent and reliable recovery of high quality total RNA that is appropriate for amplification-based methodologies.
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Figure 3. Yield from several cell lines using 1 x
105 cells is consistently 0.3µg or more when using the SV 96 Total RNA Isolation System. Purity is exceptional, with
A260/A280 ratios of 1.9 or higher. |
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Table 1. Average Yield and Purity of Total RNA Purified from 1 x
105 HeLa, NIH3T3 and CHO Cells.
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| Cell Type |
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Average Yield (µg) |
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Average A260/A280 |
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| HeLa |
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0.45 ± 0.08 |
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2.0 ± 0.1 |
| NIH3T3 |
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0.36 ± 0.09 |
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1.9 ± 0.2 |
| CHO |
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0.33 ± 0.08 |
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1.9 ± 0.2 |
High Purity - Total RNA is free of detectable genomic DNA.
The SV 96 Total RNA Isolation System incorporates a DNase treatment step designed to substantially reduce genomic DNA contamination and therefore improve RT-PCR analysis of purified total RNA.
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Figure 4. RT-PCR amplification from purified total RNA with or without DNase treatment from mouse liver lysate. Amplification primers used for the RT-PCR were designed to amplify both mRNA and DNA in the same amplification reaction. Amplification products specific for mRNA are 400bp. Amplification products specific for genomic DNA are 1.2kb. |
Additional Information
On-line Catalog
DNA and RNA Purification
Integrated Solutions in Nucleic Acid Purification Guide
For Laboratory Use
U.S. Pat. No. 6,218,531 and other patents pending
The PCR process is covered by patents issued and applicable in certain countries. Promega does not encourage or support the unauthorized or unlicensed use of the PCR process.
Biomek is a registered trademark of Beckman Coulter, Inc.
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