Name: Plant microRNA Purification Kit
Brand: Norgen Biotek Corp.
SKU: 54700
Price: 249.000000 USD
Availability: InStock

Features and Benefits

Fast and easy processing using a rapid spin-column format
No phenol or chloroform extractions
Isolate all small RNA molecules (<200 nt)
Minimal contamination from large RNA molecules and genomic DNA
High quality small RNA can be used in various downstream applications
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s Plant microRNA Purification Kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured plant cells or plant tissues. These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long, and regulate gene expression by binding to mRNA molecules and affecting their stability or translation. The small RNA molecules isolated using Norgen’s Plant microRNA Purification Kit can be used in various downstream applications relating to gene regulation and functional analysis, including RT-PCR, miRNA sequencing, northern blotting and microarray analysis.

Details

Supporting Data

Figure 1. Fractionation of Large and Small RNA. Large RNA and small RNA were sequentially purified using Norgen's Plant miRNA Purification Kit from 50 mg of raspberry leaf tissue and the RNA profile was compared with the RNA isolated using Norgen's Plant/Fungi Total RNA Purification Kit (Cat. #25800). For visualization, 10 µL of RNA from 50 µL of RNA elution was loaded on 2% 1x MOPS agarose gel. Norgen's kit was able to isolate both the large and small RNA fractions, and the small RNA fraction does not contain any of the large RNA species.

Figure 2. Efficiency of the concentration of miRNA using Norgen's Plant miRNA Purification Kit to detect three plant miRNAs. Small RNA was concentrated from 50 mg raspberry using Norgen's Plant miRNA Purification Kit. Three µL of the elution was directly used for cDNA generation and 3 µL of cDNA was used for the CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad). Blue: concentrated small RNA from Norgen's Plant miRNA Purification Kit, Red: Total RNA from Plant Total RNA Purification Kit (Cat. 25800), Circle: +RT, Cross: - RT.

Figure 3. Isolation of Concentrated RNA that can be used in Sensitive Downstream Applications. Three different miRNA purification methods were compared for their ability to isolate and detect plant miRNA. miRNA was purified from grape using three miRNA purification kits according to the manufacture’s manual. Next, 3 µL of the elution was directly used for cDNA generation and 3 µL of cDNA was used for the CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad) to detect the miRNA 159a. As it can be seen, only the miRNA purified using Norgen's kit (blue lines) was amplified, while the miRNA isolated using the other 2 kits failed to amplify (red and green lines). Both of the competitor’s kits rely on phenol to isolate the miRNA, and therefore residual phenol is likely interfering with the amplification. Norgen's kit isolates high yields of good quality miRNA without the use of phenol. Blue: Norgen Plant miRNA, Red: Competitor Q, Trizol: Green.

Kit Specifications
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Minimum Elution Volume	20 μL
Size of RNA Purified	< 200 nt
Amount of Starting Material: Plant Tissues	100 mg 5 x 10⁶ cells
Time to Complete 10 Purifications	25 minutes

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 54700 (25 preps)
Lysis Buffer C	30 mL
Wash Solution A	18 mL
Elution Solution A	6 mL
Filter Columns	25
Large RNA Removal Columns	25
microRNA Enrichment Columns	25
Collection Tubes	75
Elution Tubes (1.7 mL)	50
Product Insert	1

Citations

Title	Comprehensive analysis of both long and short read transcriptomes of a clonal and a seed-propagated model species reveal the prerequisites for transcriptional activation of autonomous and non-autonomous transposons in plants
Journal	Mobile DNA. 2022.
Authors	Chen TH, Winefield C

Plant microRNA Purification Kit