The NucleoSpin RNA Stool kit is designed for the efficient isolation of RNA, including small RNA species, from fresh and frozen stool samples. The kit combines a chemical disruption of the stool sample using NucleoZol and Lysis Buffer RST1 with a mechanical lysis using MN Bead Tubes Type A (which contain ceramic beads). No protease digestion is required.
Combination of mechanical homogenization and chemical lysis ensures high RNA recovery, including small RNA
Effective removal of PCR inhibitors
Suitable for various human and animal stool samples
Silica membrane technology combined with MN Bead Tubes Type A
Format
Mini spin columns
Sample material
Fresh or frozen stool samples
Fragment size
≥18 nt
Typical yield
10–30 µg
A260/A280
1.9–2.1
A260/A230
1.8–2.5
Typical RIN (RNA integrity number)
>7.5
Elution volume
100 µl
Preparation time
70 min/10 preps
Binding capacity
200 µg
Figure 1. The NucleoSpin RNA Stool procedure.
Bead tube disruption
Figure 2. The NucleoSpin RNA Stool kit includes MN Bead Tubes Type A. The NucleoSpin RNA Stool kit utilizes MN Bead Tubes Type A (Panel A), 0.6–0.8-mm ceramic beads included in the kit, for efficient mechanical disruption of stool samples. The beads are designed to be used in conjunction with the MN Bead Tube Holder (Cat. No. 740469) on a Vortex-Genie 2 (Panel B) or with a mixer mill (see below). MN Bead Tubes Type A are also available separately.
High yields
Figure 3. NucleoSpin RNA Stool provides higher yields than competitor kits. Two human stool samples were processed in triplicates using 250 mg of stool sample with two competitor kits (Q and Z) and only 200 mg of stool sample with NucleoSpin (MN), all according to their standard protocols. The Competitor Q protocol that was performed included an optional phenol-based lysis step. Each eluate was analyzed using UV spectroscopy to determine RNA yield. The samples processed using NucleoSpin RNA Stool showed the highest RNA yield of all the human stool samples.
qRT-PCR performance
Figure 4. NucleoSpin RNA Stool provides better qPCR performance than competitor kits. The NucleoSpin RNA Stool samples and the Q samples were each eluted in 100 μl, while the Z eluates were adjusted to 100 μl to obtain comparable RNA concentrations. 1 μl of eluate was used for qRT-PCR of a 466-bp amplicon on a Roche LightCycler using a one-step qRT-PCR kit. NucleoSpin RNA Stool (MN) showed better qRT-PCR performance than the two competitor kits (Q and Z).