Circulating RNA purification from plasma/serum samples

The Plasma/Serum Circulating and Exosomal RNA Purification Kits (Slurry Format) from Norgen allow for the quick and easy extraction of circulating and exosomal RNA from plasma/serum samples.

Format

Mini slurry

Size

50 Preps

Image Credit: Norgen Biotek Corp.

Slurry

Size

50 preps

Image Credit: Norgen Biotek Corp.

High throughput slurry

Size

1 × 96-Well Plate

Image Credit: Norgen Biotek Corp.

Maxi slurry

Size

25 Preps

Image Credit: Norgen Biotek Corp.

Overview

Norgen's kits can isolate all types of circulating and exosomal RNA, including microRNA, without the need for phenol or chloroform. The slurry format has an advantage over other kits because it does not require extension tubes to purify free-circulating and exosomal RNA from large sample volumes. This kit can isolate RNA from both fresh and frozen materials. This kit is designed to isolate RNA from serum or plasma prepared from blood collected only on EDTA or citrate.

Plasma samples prepared from blood collected with heparin should be avoided, as heparin can significantly interfere with many downstream applications, including RT-PCR. The purified plasma/serum free-circulating and exosomal RNA is eluted in a solution that is fully compatible with a range of applications, such as PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection assays, primer extension, expression arrays, and NGS.

Free-circulating RNA in plasma and serum can serve as tumor- and fetal-specific markers, making them valuable for both cancer detection and prenatal diagnostics. In addition, these RNAs hold promise as biomarkers for a range of other disease states. Typically, free-circulating RNA in plasma or serum exists as short fragments of less than 1000 nucleotides, while free-circulating microRNAs (approximately 21 nucleotides) are also commonly detected in these fluids.

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Features and benefits

• Available in Spin Column or 96-well plate formats
• Bind and elute all RNA, irrespective of size or GC content, without bias
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Concentrate circulating and exosomal RNA into small elution volumes
• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Isolate inhibitor-free circulating and exosomal RNA
• Process 96-well plates using vacuum or centrifugation
• Purification is based on Norgen’s proprietary resin matrix
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing.
• Versatile plasma and serum input volumes

Plasma/serum circulating and exosomal RNA purification kit (Slurry)

Norgen's Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) is a reliable, fast, and simple method when it comes to isolating circulating RNA and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 5 mL.

Plasma/serum circulating and exosomal RNA purification mini kit (Mini slurry)

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Slurry Format) is a quick, dependable, and easy method for separating circulating and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 2 mL.

Plasma/serum circulating and exosomal RNA purification 96-well kit (High throughput slurry)

Norgen's Plasma/Serum Circulating and Exosomal RNA Purification 96-Well Kit (Slurry Format) uses a vacuum manifold or centrifugation to isolate circulating and exosomal RNA from plasma and serum samples in a high-throughput manner.

Plasma/serum circulating and exosomal RNA purification maxi kit (Maxi slurry)

Norgen's Plasma/Serum Circulating and Exosomal RNA Purification Maxi Kit (Slurry Format) offers a quick, dependable, and easy method for separating circulating RNA and exosomal RNA from different volumes of plasma/serum, ranging from 2 mL to 5 mL.

Details

Supporting data

Figure 1. Isolation and Detection of Circulating RNA from Different Plasma Volumes. Norgen's Plasma/Serum Circulating RNA Purification Mini Kit (Slurry Format) was used to isolate circulating RNA from 0.5 mL, 1 mL and 2 mL plasma. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene. The 5S housekeeping gene was detected from all plasma sample volumes used. The amplification of the 5S rRNA showed an increasing amount of RNA with increasing the sample input volume. This is represented by the decrease of the Ct value with increasing the sample input volume. RNA isolated from 0.5 mL plasma is represented by the red line, RNA isolated from 1 mL plasma is represented by the green line, and RNA isolated from 2 mL plasma is represented by the blue line. The black line corresponds to the no template control. Image Credit: Norgen Biotek Corp.

Figure 2. Effective Isolation of Plasma Circulating RNA from Different Volumes. Norgen's Plasma/Serum Circulating RNA Purification Mini Kit (Slurry Format) was used to isolate circulating RNA from 0.5 mL, 1 mL and 2 mL plasma. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene. The 5S housekeeping gene was detected from all plasma sample volumes used. The amplification of the 5S rRNA showed an increasing amount of RNA with increasing the sample input volume. This is represented by the decrease of the Ct value with increasing the sample input volume. Average Ct. values for the amplification of the 5S rRNA isolated from 0.5 mL plasma is represented by the Red bar, Average Ct. values for the amplification of the 5S rRNA isolated from 1 mL plasma is represented by the Green bar, whereas Average Ct. values for the amplification of the 5S rRNA isolated from 2 mL plasma is represented by the blue bar. Image Credit: Norgen Biotek Corp.

Figure 3. Effective and Consistent Detection of Plasma Exosome RNA. Norgen's Plasma/Serum Circulating and Exosomal RNA Isolation Kits can effectively isolate RNA from plasma. Plasma Exosome RNA was isolated from 500 µL of human plasma prepared from blood collected on citrate, EDTA or Heparin in triplicates using Norgen's Plasma/Serum RNA Isolation Kit (blue), a ExoQuick Exosome Precipitation Reagent (green) and Qiagen’s QIAzol extraction followed by a modified Qiagen's RNeasy Mini Kit cleanup (red). Stem loop RT-qPCR using primers specific to miR-21 and miR-16 as well as the housekeeping 5S rRNA was performed. In brief, three microliters of the 100 µL isolated RNA was then subjected to a 20 µL reverse transcription using 5S rRNA, miR-21 and miR-16 stem-loop reverse primer or reveres primer. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21, human miR-16 and the 5S rRNA. Norgen's Plasma/Serum Exosome RNA Isolation Kit is the only product that showed consistent detection of all tested transcripts with the highest quality regardless the type of the anti-coagulant used for blood collection. Image Credit: Norgen Biotek Corp.

Figure 4. Isolation and Detection of Circulating RNA from Different Plasma Volumes. Norgen's Plasma/Serum Circulating RNA Purification Maxi Kit (Slurry Format) was used to isolate circulating RNA from 2 mL, 3 mL and 5mL plasma. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene. The 5S housekeeping gene was detected from all plasma sample volumes used. The amplification of the 5S rRNA showed an increasing amount of RNA with increasing the sample input volume. This is represented by the decrease of the Ct value with increasing the sample input volume. RNA isolated from 2 mL plasma is represented by the Red line, RNA isolated from 3 mL plasma is represented by the Green line whereas RNA isolated from 5 mL plasma is represented by the Blue line. The black line corresponds to the no template control. Image Credit: Norgen Biotek Corp.

Figure 5. Effective Isolation of Plasma Circulating RNA from Different Volumes. Norgen's Plasma/Serum Circulating RNA Purification Maxi Kit (Slurry Format) was used to isolate circulating RNA from 2 mL, 3 mL and 5 mL plasma. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene. The 5S housekeeping gene was detected from all plasma sample volumes used. The amplification of the 5S rRNA showed an increasing amount of RNA with increasing the sample input volume. This is represented by the decrease of the Ct value with increasing the sample input volume. Average Ct. values for the amplification of the 5S rRNA isolated from 2 mL plasma is represented by the Red bar, Average Ct. values for the amplification of the 5S rRNA isolated from 3 mL plasma is represented by the Green bar whereas Average Ct. values for the amplification of the 5S rRNA isolated from 5 mL plasma is represented by the blue bar. Image Credit: Norgen Biotek Corp.

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Mini slurry

Source: Norgen Biotek Corp.

Cat.51000 Storage Conditions and Product Stability: The Slurry C1 and Lysis Buffer A should be warmed up for 20 minutes at 60 °C if any salt precipitation (crystallization) is observed. Slurry C1 contains grey resin that will not disappear by warming up.

Slurry

Source: Norgen Biotek Corp.

Storage Conditions and Product Stability: All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. If any salt precipitation is observed, it is recommended to warm Slurry C2 and Lysis Buffer A for 20 minutes at 60 °C. Slurry C2 contains a grey resin that will not fade with warming up.

High throughput slurry

Source: Norgen Biotek Corp.

Cat.29500 Storage Conditions and Product Stability: All solutions should be stored at room temperature and kept tightly sealed. This kit is stable for two years following the date of shipment.

Maxi slurry

Source: Norgen Biotek Corp.

Storage Conditions and Product Stability: All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for two years following the shipment date.

Source: Norgen Biotek Corp.