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.

Description

The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.

Features

• 5’→3’ DNA polymerase activity
• No detectable 3’→5′ exonuclease (proofreading) activity
• Generates PCR products with 3′-dA overhangs
• High yield PCR 
• High reproducibility
• Reduced  pipetting errors
• For direct loading into electrophoresis analysis
• DNA bands can be visualized directly under UV or 470 nm blue light illumination

Applications 

• Routine PCR
• Colony PCR
• High throughput PCR
• Amplification of DNA fragments up to 8 kb
• Generation of PCR products for TA cloning
• DNA labeling

Storage

Protected from light and avoid multiple freeze/thaw cycles
4°C  for     6 months
-20°C  for  24 months

The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.

Introduction

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.

Details

Specifications

Principle

The HiPure silica gel column uses a high binding ability glass fiber filter membrane as the substrate. Under the condition of high concentration of ionizing agent (such as Guanidinium chloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bonding and electrostatic and other physical factors, while protein or other impurities are not adsorbed and removed. The filter membrane that has adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (such as Buffer TE) or water can be used to wash out the nucleic acids adsorbed on the filter membrane. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

The stool samples are homogenized in the lysis solution, further lysed in a high-temperature water bath, and RNA is released into the lysis solution. Chloroform extraction removes genomic DNA and impurities, transfer the supernatant to an alcohol free binding solution, purify RNA through a column, and finally elute RNA with RNase Free Water. The purified RNA can be directly used for experiments such as PCR, Southern hybridization, and enzyme digestion.

Advantages

• High purity – unique adsorbent for more efficient removal of inhibitors
• High concentration – maximum extraction of total RNA from stool samples
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions. At low temperatures, Buffer SPL may form precipitates, dissolve it by 55°C water bath. After receiving the product, Buffer PHC should be stored at 2-8°C.

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.