Catalog No. : MN-740903.50
NucleoSpin RNA Clean-up XS (50)
- Complete removal of RT-PCR inhibitors
- Very high RNA recovery and concentration
- RNA clean-up from pre-purified RNA (phenol / chloroform), enzymatic reaction (e.g., amplification reactions, labeling reactions, DNase digestions)
- Time-saving procedure based on NucleoSpin® RNA XS technology
- Input volume up to 300 μL
- Elution in as little as 5 μL
|NucleoSpin RNA Clean-up XS (50)||
NucleoSpin® RNA Clean-up XS Columns with Collection Tubes, Collection Tubes (2 mL), Collection Tubes (1.5 mL), buffers
The NucleoSpin® RNA Clean-up XS kit is designed for clean-up and concentration of pre-purified RNA samples. Typical sample material covers picogramm to microgramm amounts of pre-purified RNA (e.g., phenol-purified RNA) and RNA from reaction mixtures (e.g., DNase treated samples).
The innovative column design with a funnel shaped thrust ring and a small silica membrane area allows processing of sample volumes of up to 300 µL and elution of RNA in as little as 5–30 µL. Thus, highly concentrated RNA is eluted, ready-to-use for sensitive downstream applications (e.g., RT-PCR). RNA enrichment of 20x up to 50x can be achieved. The RNA recovery is typically about 85–95%.
High quality RNA (RNA Integrity Number (RIN) > 9 according to Agilent 2100 Bioanylzer assays) can be obtained from high quality RNA samples. The RIN of the processed sample is typically equal (±0.3) to the RIN of the input sample.
The NucleoSpin® RNA Clean-up XS kit allows clean-up and concentration of RNA with an A260/A280 ratio generally exceeding 1.9. Due to the high RNA purity large amounts of eluates can successfully be used as template in RT-PCR without inhibition.
NucleoSpin® RNA Clean-up XS procedure
The innovative funnel shaped thrust ring of the NucleoSpin® RNA XS Column holds a silica membrane of very small diameter.
Thus very efficient elution in only 5–30 μL can be achieved resulting in up to 50x concentrated RNA.
Efficient removal of contaminants
RNA was isolated from mouse liver with TRIzol®. 100 µL crude extract were subjected to the NucleoSpin® RNA Clean-up XS procedure. Some of the samples were treated with 10 µL of a rDNase/Reaction Buffer mixture and incubated for 10 minutes before clean-up.
The A260 nm/A280 nm ratio (A260/A280) was measured for eight samples, respectively. Optimal A260/A280 values for RNA solutions are 1.9–2.0.
Crude TRIzol® extracts show poor results due to contaminations (e.g., phenol). Samples, purified with NucleoSpin® RNA Clean-up XS have an average ratio A260/A280 of 1.9.
NucleoSpin® RNA Clean-up XS efficiently removes contaminants, resulting in pure RNA. The rDNase digestion enables removal of contaminating DNA without affecting purity.
Purification of high-quality RNA from reaction mixtures
A 100 µL crude RNA sample (contaminated with DNA) was treated with 10 µL MACHEREY-NAGEL rDNase/Reaction Buffer mixture, incubated for 10 min and subjected to the NucleoSpin® RNA Clean-up XS procedure. The RNA was finally eluted in 20 µL. RNA quality (RIN) and quantity were analyzed with an Agilent Bioanalyzer, Agilent RNA 6000 Nano chip, and Agilent RNA 6000 Nano reagent before and after clean-up. The RIN (RNA integrity number) is an indicator for RNA quality (the optimal RIN value for non degraded RNA is 10).
The crude RNA solution (lane 1) shows a low RNA concentration of 85 ng/µL. Nevertheless, the RNA is not degraded and has a high RIN of 9.4.
Lane 3 shows the same RNA after an enzymatic reaction (DNase digestion) and purification with NucleoSpin® RNA Clean-up XS.
RNA quality is not affected by the whole procedure, the RIN of 9.7 is as high as before clean-up. The high recovery of 95% leads to a concentration of 404 ng/µL in a total volume of 20 µL.
Lane L: RNA Ladder
Lane 1: Crude RNA solution (100 μL total volume): concentration 85 ng/μL, RIN 9.4. Due to the low concentration of the crude RNA solution, the brightness of the lanes are individually adjusted.
Lane 3: NucleoSpin® RNA Clean-up XS eluate (20 μL total volume): concentration 404 ng/μL, RIN 9.7
NucleoSpin® RNA Clean-up XS is ideal for purification of RNA from reaction mixtures. The eluted RNA is of best possible quality and RIN, with a recovery of 85–95%.
High concentration of dilute RNA solutions
Details: An RNA solution with a concentration of 10 ng/µL was subjected to NucleoSpin® RNA Clean-up XS and to a competitor RNA Clean-up kit (MinElute).
In order to get the best possible concentration of RNA, highest possible volumes of the RNA solution were loaded on the columns and elution was performed with the lowest possible volumes, specified in the kit protocols.
A) NucleoSpin® RNA Clean-up XS: 300 µL RNA solution – elution volume 5 µL
B) Competitor kit, RNA Cleanup - MinElute: 200 µL RNA solution – elution volume 10 µL
RNA concentrations of the eluates were measured with RiboGreen fluorescent dye before and after clean-up. NucleoSpin® RNA Clean-up XS shows the best results. The concentration factor is 51x which is more than three times higher than the concentration factor of the competitor kit (16x).
Highest RNA concentration was achieved with MACHEREY-NAGEL’s NucleoSpin® RNA Clean up XS. It allows concentrating RNA more than 50 fold, a three times higher factor than the competitor kit.