|Minimum Plasma/Serum Input||50 μL|
|Maximum Plasma/Serum Input||200 μL|
|Time to Complete Purification||15-20 minutes|
|Size of RNA Purified||All sizes, including miRNA and small RNA (< 200 nt)|
|Plasma/Serum RNA Purification Kit Benefits|
|No phenol:chloroform extractions||Circulating RNA and Exosomal RNA is isolated without the use of harmful chemicals such as phenol or chloroform|
|Isolate all sizes of circulating RNA and exosomal RNA||The kit allows for the isolation of all sizes of fragmented circulating RNA and exosomal RNA, including microRNA|
|Fast and easy processing||Rapid spin column format allows for the processing of multiple sam-ples in under 15-20 minutes.|
|Small input volume||Isolate circulating RNA and exosomal RNA from 50 μL to 200 μL of plasma/serum.|
|Concentrate circulating RNA and Exosomal RNA||Circulating and exosomal RNA present in input volumes of 50 μL to 200 μL are concentrated into final elution volume of 10 μL to 25 μL.|
|Isolate inhibitor-free RNA||Purified RNA can be used in a number of sensitive downstream ap-plications including reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays|
The purified circulating RNA is compatible with most of the molecular biological applications such as:
- Reverse transcription qPCR
- Reverse transcription PCR
- Northern blotting
- RNase protection and primer extension
- Expression array assays.
- miRNA array
- Droplet/Digital PCR
Figure 1. Effective and Consistent Detection of miRNA from Plasma. Norgen's Plasma/Serum RNA Purification Kit can effectively isolate miRNA from plasma. Circulating miRNA was isolated from 200 µL plasma using Norgen's Plasma/Serum RNA Purification Kit, Company A's kit and Company C's kit. Circulating miRNA was isolated from 600 µL using Company B's Kit. Stem loop RT-qPCR using primers specific to miR-21 was performed. In brief, 1 microliter of the 15 µL purified RNA using Norgen's Plasma/Serum RNA Purification Kit, Company A's kit and 3.3 microliters of the 50 µL purified RNA using Company C's kit and Company B's kit was then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse 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. Norgen's Plasma/Serum RNA Purification Kit is the only product that showed the most consistent and the highest recovery of the miR-21 transcripts as compared to the other isolation methods. The recovery of the miRNA from 200µL plasma was higher than that recovered from RNA purified from 600µL using Company B's kit.
Figure 2. Purification of Circulating RNA from Different Plasma Volumes. Norgen’s Plasma/Serum RNA Purification Kit was used to purify circulating RNA from 50µL, 100µL and 200µL plasma prepared from blood collected on EDTA. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect miR-21 (Figure 1A) and the housekeeping 5S rRNA transcript (Figure 1B). The relative amount of both the miR-21 (Figure 1A) and the 5S rRNA transcript (Figure 1B) is linearly increasing with increasing the sample input volume.
Figure 3. Eluting Purified Circulating RNA from into Different Elution Volumes. Norgen’s Plasma/Serum RNA Purification Kit was used to purify circulating RNA from 200µL plasma prepared from blood collected on EDTA and eluted in 10µL, 15µL, and 25µL. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect miR-21 (Figure 2A) and the housekeeping 5S rRNA transcript (Figure 2B). The relative amount of both the miR-21 (Figure 1A) and the 5S rRNA transcript (Figure 1B) is decreasing with increasing the elution volume indicating the efficient concentration of the plasma circulating RNA in a very low elution volume.
|Item||Cat. No.||Kit Size|
|Plasma/Serum RNA Purification Kit||55000||50 preps|
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