SNPsig® SARS-CoV-2 (E484K) Identification Kit

Overview

• Well-accepted microRNA sequence used for normalization in gene expression studies
• Best suited for RNA purification from samples with low RNA abundance including liquid biopsies (plasma/serum/urine etc.) and low cell or tissue inputs
• Compatible to expression analysis using RT-qPCR – both RNA and matching forward PCR primer provided.
• Fully compatible to Norgen’s microScript cDNA Synthesis system
• Fully compatible to Next Generation Sequencing (Small RNA-Seq) library preparation workflow

The amount of RNA that can be extracted from different biological or clinical samples varies greatly. For example, while a few micrograms of RNA could be easily purified from tissues and cells in excess amounts (such as from a few milligrams of tissue), many liquid biopsy samples may yield very low amounts of RNA. In fact, samples such as urine or plasma may yield 1 – 100 ng or less RNA per 100 µL of sample. Such a range of RNA quantity is often below the detection limit of most commonly used techniques for measuring RNA including nano-spectrophotometry and fluorescent nucleic acid stains. As a result, without properly determined RNA concentration, it becomes very difficult to normalize the starting quantity of RNA used in gene expression studies.

Norgen’s microRNA (cel-miR-39) Spike-In Kit offers a quantified synthetic RNA (cel-miR-39) for spike-in during RNA extraction procedures and subsequent normalization in RT-qPCR assays. The amount of cel-miR-39 RNA recovered after RNA extraction is directly correlated with the amount of total RNA recovered. After reverse transcription (such as with Norgen’s microScript Reverse Transcription system) of the sample RNA (with spike-in), the level of cel-miR-39 can be determined by subjecting the cDNA generated to quantitative PCR (qPCR) using fluorescent nucleic acid stains such as SYBR Green.  A cel-miR-39 specific primer is included in the kit. The level of expression of any target transcripts in different RNA samples can now be normalized to the cel-miR-39 transcript level using standard method such as ∆∆Ct relative quantification.

In addition, the cel-miR-39 RNA is compatible to library preparation methods (including ligation-based protocols) in Next Generation Sequencing (Small RNA-Seq) workflows. The cel-miR-39 RNA could be used for normalization as well as for tracking library construction efficiency.

Details

Supporting Data

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Storage Conditions
Upon receipt, store Norgen’s microRNA (cel-miR-39) Spike-In Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• ​Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment, nor carrier RNA
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents, nor overnight incubation required
• Concentrate isolated exosomal RNA and are free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL.

Details

Supporting Data

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*Please check page 5 of the product insert for the average yields and the common RNA quantification methods

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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Product Description

RNA Amplification Kit With Improved Detection & Quantification-20°C Format Easy And Efficient

RNA Amplification Kit With Improved Detection & Quantification-20°C Format Easy And Efficient

Product Detail

Kit Storage and term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal nucleic acid Principle Summary

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39 ºC~42 ºC), reverse transcriptase uses specific primers and template RNA to synthesize cDNA strands, and binds the auxiliary protein and single strand With the help of the protein, the recombinase and the primer form a complex; perform a homology search and bind the target homology domain, at this time a D-loop region is formed at the homology position and strand exchange begins; accompanied by the recombinase from the complex Upon dissociation, the polymerase also binds to the 3′ end of the primer, initiating chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.

Isothermal nucleic acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.

Technical Parameters:

Isothermal nucleic acid Applications

Suitable for RNA isothermal rapid amplification kit(NFO type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

RNA NFO kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.