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Total RNA Purification

Eliminate Genomic DNA Contamination with NucleoSpin RNA II

Total RNA Isolation with Genomic DNA Removal You Can See!

While the NucleoSpin RNA II Kit and Vendor Q’s RNeasy Mini kit provide similar yields of total RNA, Competitor Q’s RNA isolation kit leaves genomic DNA contamination.

NucleoSpin RNA isolation kit removes gDNA; RNeasy does not.

RNA was purified using the NucleoSpin RNA II Kit or Competitor Q’s RNeasy Mini kit. 10 µl of purified RNA was treated with RNase A and loaded onto a 1% TAE agarose gel to reveal any contaminating genomic DNA. Genomic DNA was completely eliminated from samples purified with the NucleoSpin RNA II kit, but RNA purified with the RNeasy Mini kit showed evidence of genomic DNA contamination.

NucleoSpin RNA II gives pure total RNA; no genomic DNA.

RNA was purified using the NucleoSpin RNA II Kit or Competitor Q’s RNeasy Mini kit. 10 µl of purified RNA was treated with RNase A and loaded onto a 1% TAE agarose gel to reveal any contaminating genomic DNA. Genomic DNA was completely eliminated from samples purified with the NucleoSpin RNA II kit, but RNA purified with the RNeasy Mini kit showed evidence of genomic DNA contamination.

 

How Does NucleoSpin RNA II Remove Genomic DNA Contamination?

If you want to isolate high-quality RNA, it is important to prevent degradation of the RNA and to eliminate genomic DNA. Unlike Competitor Q’s RNeasy Kits, NucleoSpin RNA II includes the following:

  • An optimized DNase I treatment to eliminate genomic DNA contamination
  • Filters/shredders to homogenize samples and reduce viscosity

Without these components, RNA purified with Competitor Q’s RNeasy kits exhibits significant DNA contamination.

Total RNA Purification from Just a Few Cells

With NucleoSpin RNA II, you can detect transcripts from very small numbers of cells—in this example, RNA was isolated from as few as 10 HeLa cells.

NucleoSpin RNA II can isolate total RNA from 10 cells.

Total RNA was isolated from the indicated number of HeLa cells with NucleoSpin RNA II (elution volume: 40 µl). RT-PCR reactions were carried out with 3 µl of total RNA and beta-actin primers. The RT-PCR product was 626 bp.

Simple Protocol for DNA Removal

NucleoSpin RNA II uses a simple spin-column procedure that is probably already familiar to you. First, cells are lysed in a buffer that inactivates RNases (which are present in virtually any biological material) and creates appropriate conditions for RNA to bind to the silica membrane. After lysis, homogenization and reduction of viscosity are achieved by filtration with NucleoSpin Filter units (shredders), which are provided with the kit. Residual genomic DNA is removed by on-column digestion with rDNase, which is also included in the kit.

NucleoSpin RNA II: simple RNA purification protocol.

What is NucleoSpin RNA II?

Technology Silica membrane
Format Mini spin columns
Starting material 5 x 106 cultured cells or 109 bacterial cells or 30 mg tissue
Fragment size >200 bases
Yield 14–70 µg total RNA
Elution volume 40–100 µl
Time 30 min/6 preps

 
 
Products
Cat. # Product Package Size Price # of Units Select
740955.250 NucleoSpin® RNA 250 Preps $1,072.00
740955.50 NucleoSpin® RNA 50 Preps $233.00
740955.10 NucleoSpin® RNA (Sample-Not for Sale) 10 Preps $51.00
 

 

 

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