Dengue Virus

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.

Component	Cat. 59000
cel-miR-39 RNA	10 pmol
cel-miR-39 Forward PCR Primer	1 nmol
Nuclease-Free Water	1.25 mL
Product Insert	1

Overview

• Ready-to-use
• Quantitative
• Highly Stable
• Precise
• Eight discrete fragments from 50 bp to 2000 bp
• Higher intensity reference band at 500 bp

The FastRunner DNA Ladder is prepared to ensure quality and batch-to-batch consistency. This Ladder contains eight discrete fragments ranging from 50 bp to 2000 bp with a higher intensity reference band at 500 bp. This Ladder is ideal for quick sizing of PCR products and restriction digests.

Contents
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).

Details

Supporting Data

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FastRunner DNA Ladder (Cat# 12800) – 100 loads

Ladder Properties:
• Eight discrete bands, ranging from 50 bp to 2000 bp
• Higher intensity band at 500 bp for easy reference

Fragment	Size (bp)	Mass (ng)
1	2000	104
2	1500	88
3	1000	68
4	750	59
5	500	93
6	300	28
7	150	35
8	50	25

Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.

Storage: 
Stable at room temperature. For longer term storage, -20°C is recommended.

This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.