Norgen’s microScript microRNA cDNA Synthesis Kit is an all-in-one, ready-to-use product for the reverse transcription of microRNA from either Total RNA preparations or enriched microRNA preparations. The kit contains the 2x Reaction Mix and the microScript microRNA Enzyme Mix. The kit utilizes Norgen’s microScript Reverse Transcriptase, a mutant version of Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase. It has reduced RNase H activity and increased thermal stability.
The workflow of Norgen’s microScript microRNA cDNA Synthesis Kit involves a simple, single-tube set-up by the mixing of 2x Reaction Mix, Enzyme Mix and the RNA template. The reaction can then be carried out in a thermocycler. A poly (A) tail is first added to the RNA template, followed by cDNA synthesis using an adapter primer. In addition to the ease-of-use, the single-tube set-up provides superb consistency and sensitivity. The cDNA could be used in a PCR or qPCR amplification using a Universal PCR Reverse Primer and the forward primer that contains the sequence of the microRNA of interest. A single cDNA preparation could be used for PCR amplification of a number of different microRNAs. In addition, the cDNA preparation could be used for PCR or qPCR detection (using gene-specific forward and reverse primers) of mRNA or large RNA if total RNA preparation was the starting template. This could allow for parallel evaluation of expression level of microRNAs and microRNA-targets.
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Storage Conditions and Product Stability
This kit is stable for 1 year after the date of shipment .
| Component | Cat. 54415 (12 rxns) | Cat. 54410 (50 rxns) |
|---|---|---|
| microScript microRNA Enzyme Mix | 12 μL | 50 μL |
| 2x Reaction Mix | 120 μL | 500 μL |
| Universal PCR Reverse Primer | 60 μL | 250 μL |
| Nuclease-Free Water | 1.25 mL | 2 x 1.25 mL |
| Product Insert | 1 | 1 |
Propargyl-PEG12-methylamine is a propargyl linker that is commonly used as a Click Chemistry reagent for copper-catalyzed reactions with azides. The methylamine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. The PEG spacer helps improve the water solubility of the molecule in aqueous media. Reagent grade, for research use only.
Propargyl-PEG12-methylamine is a propargyl linker that is commonly used as a Click Chemistry reagent for copper-catalyzed reactions with azides. The methylamine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. The PEG spacer helps improve the water solubility of the molecule in aqueous media. Reagent grade, for research use only.
Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.
This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.
Urine DNA Isolation Kit
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.
Urine DNA Isolation Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.
Urine DNA Isolation Maxi Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.
Background
DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.
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| Kit Specifications – Spin Column | |
| Minimum Urine Input | 3 mL |
| Maximum Urine Input | 25 mL |
| Time to Complete 10 Purifications | < 30 minutes |
| Size of Urine DNA Purified | Large (> 1 kb) and small (150-250 bp) |
| Kit Specifications – 96-well | |
| Urine Input Volume | 3 – 25 mL |
| Elution Volume | 1st Elution 100 μL 2nd Elution 100 μL |
| Size of DNA Purified | Large (> 1 kb) and small (150-250 bp) |
| Time to Complete 96 Purifications | < 1 hour |
Storage Conditions and Product Stability
All buffers 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 up Slurry B1 and Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed. Slurry B1 contains a grey resin that will not dissolve when warmed.
| Component | Cat. 18100 (50 preps) | Cat. 48800 (50 preps) | Cat. 50100 (50 preps) |
|---|---|---|---|
| Binding Solution K | 15 mL | – | – |
| Slurry B1 | – | 18 mL | 18 mL |
| Proteinase K in Storage Buffer | 2 mL | – | – |
| Pronase K in Storage Buffer | 2 mL | – | – |
| Soluton WN | 9 mL | – | – |
| Binding Buffer A | – | – | 50 mL |
| Lysis Buffer A | – | 30 mL | 30 mL |
| Wash Solution A | – | 38 mL | 38 mL |
| Wash Solution B | 30 mL | – | – |
| Wash Solution D | 9 mL | – | – |
| Binding Solution K | 15 mL | – | – |
| Elution Buffer B | 15 mL | 15 mL | 15 mL |
| Micro Spin Columns | 50 | – | – |
| Mini Filter Spin Columns | – | 50 | 50 |
| Collection Tubes | 50 | 50 | 50 |
| Elution Tubes (1.7 mL) | 100 | 100 | 100 |
| Product Insert | 1 | 1 | 1 |