NGS Library Quantification Kit (for Small RNA-Seq)
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Detail
Overview
Able to quantify NGS Library (Illumina) of a wide spectrum of concentrations, including sub-nanomolar concentrations
DNA is accurately quantified by using a standard curve constructed from the provided DNA Standards
Specially designed DNA standards for Small RNA-Seq library; also compatible to NGS library of other molecular weights
Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) offers a PCR-based detection procedure to quantify NGS libraries (specifically Small RNA-Seq) of a wide spectrum of concentrations. The kit consists of a specially designed primer mix compatible with the Illumina system, that is used in conjunction with the provided 2X Real-Time PCR Master Mix to amplify a library of unknown concentration. The unknown library is accurately quantified by using a standard curve constructed from the provided DNA Standard (range from 20 pM to 2 fM) on a Real-Time PCR System. The kit is specially optimized to quantify Small RNA-Seq libraries with DNA standards that have similar size to a Small RNA-Seq library. However, it could also be used with other types of NGS libraries.
Storage Conditions Upon receipt, store Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
* Quantified Library Standards are provided as 20 pM, 2 pM, 200 fM, 20 fM and 2 fM
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Product Info
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Propargyl-PEG8-amine is a heterobifunctional reagent consisting of a propargyl group and an amine group. The amine group can form amide bonds with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can form triazole linkage with azides in copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Background
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
EXTRAClean Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
EXTRAClean Plasma/Serum RNA Purification Midi Kit
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
EXTRAClean Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
All sizes, including miRNA and small RNA (<200 nt)
Average Yields¥
Variable depending on specimen
† This kit is suitable for the isolation of RNA 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
¥ Please check page 6 for Average Plasma/Serum Yields and 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.
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from <25 mg tissue, culture cells, FTA card
Applications
PCR, southern bolt and virus detection, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Animal tissue or cultured cells
Sample amount
Animal tissue : <25mg, Cultured cells : <5 x 106
Elution volume
≥30μl
Time per run
30 – 60 minutes
Liquid carrying volume per column
800μl
Binding yield of column
100μg
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while proteinis not adsorbed and is removed with filtration. After washing proteins andother impurities, Nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA).
Advantages
Good repeatability – suitable for extracting high-yield DNA from different types of tissue samples
High purity – can be used for downstream applications such as multiplex and quantitative PCR
Fast – simplified process, extracting several samples in 20 minutes (after digestion)
Safety – no phenol orchloroform extraction, no alcohol precipitation
Kit Contents
Contents
D312102
D312103
Purification Times
50 Preps
250 Preps
Buffer ATL
15 ml
65 ml
Buffer DL
15 ml
65 ml
Buffer GW1
22 ml
110 ml
Buffer GW2
12 ml
50 ml
RNase A
10 mg
50 mg
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
5 ml
15 ml
Buffer AE
15 ml
60 ml
HiPure gDNA Mini Columns
50
2 x 125
2 ml Collection Tubes
100
5 x 100
Storage and Stability
RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Purchase Guide
Document
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.
