Simultaneously clean-up and normalize PCR products
Fast (less than 20 minutes), high-throughput and easy processing using centrifugation
Sufficient elution volume (100 µL) for repeat or future assays
Non-magnetic bead purification
Norgen’s NGS Normalization 96-Well Kit allows for high-throughput NGS library PCR amplicon clean-up and normalization of NGS library PCR product concentration. The clean-up removes all PCR by-products including primers, dimers, enzymes and unincorporated nucleotides during the library prep PCR step. Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. The amount of Norgen’s matrix in each 96 well is optimized to have a limited binding capacity, thus each well can elute an equal concentration of PCR product. The process involves first adding 3 volumes of Buffer SK to the PCR product, and the mixture is then loaded onto the 96-Well Normalization Plate. The bound PCR amplicon is then washed with the provided Wash Solution A in order to remove any remaining impurities, and the purified PCR amplicon is eluted with Elution Buffer B. The purified and normalized library PCR amplicon can then be used in NGS workflows and other sequencing applications.
Storage Conditions All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component
Cat. 61900 (192 preps)
Buffer SK
30 mL
Wash Solution A
2 x 38 mL
Elution Buffer B
30 mL
96-Well Normalization Plate
2
96-Well Collection Plate
2
96-Well Elution Plate
2
Adhesive Tape
1
Product Insert
1
Other Products
Plasma/Serum RNA Purification Kits
Product Info
Document
Product Info
Overview
Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile plasma/serum input ranges
No phenol extractions
No carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Concentrate circulating RNA and exosomal RNA into a flexible elution volume
High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Compatible with Streck Cell-Free RNA BCT® Tubes
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 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.
Background
Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 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. 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. 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.
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.
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.
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.
*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 7 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.
Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 1 year under recommended storage conditions
Analyte:
D-Galacturonic Acid, D-Glucuronic Acid
Assay Format:
Spectrophotometer, Microplate, Auto-analyser
Detection Method:
Absorbance
Wavelength (nm):
340
Signal Response:
Increase
Linear Range:
5 to 150 µg of D-glucuronic acid or D-galacturonic acid per assay
Limit of Detection:
~ 15.5 mg/L
Reaction Time (min):
~ 10 min at 25oC or ~ 5 min at 37oC
Application examples:
Hydrolysates of plant material and polysaccharides and other materials.
Method recognition:
Novel method
The D-Glucuronic/D-Galacturonic test kit is a simple, reliable and accurate method for the measurement and analysis of D-hexuronic acids (specifically D-glucuronic acid and D-galacturonic acid) in plant extracts, culture media/supernatants and other materials.
Note for Content: The number of manual tests per kit can be doubled if all volumes are halved. This can be readily accommodated using the MegaQuantTM Wave Spectrophotometer (D-MQWAVE).
Mega-Calc™ software tool is available from our website for hassle-free raw data processing
Standard included
Suitable for manual, microplate and auto-analyser formats
Document
The D-Glucuronic/D-Galacturonic test kit is a simple, reliable and accurate method for the measurement and analysis of D-hexuronic acids (specifically D-glucuronic acid and D-galacturonic acid) in plant extracts, culture media/supernatants and other materials.
Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:
1. The sample is highly infectious, posing great harm to operators and the environment.
2. The source of DNA is complex and aportion of the nucleic acid, such as viral DNA or free DNA, may be lost during the operation, leading to downstream detection failure;
3. Blood sample contains a large amount of impurities and inhibitory factors.
Currently there are many methods available for extracting DNA from whole blood samples, such as phenol chloroform extraction, salting out method, etc. However, these methods require pre-treatment of blood sample, which removes red blood cells and isolate white blood cells in the first step. Due to the requirement that it cannot inactivate or kill pathogens during the process of removing red blood cells, the waste liquid (red blood cell lysate) and consumables may be contaminated by pathogens and become infectious, posing a danger to the entire laboratory environment and operators. In addition, during the process of removing red blood cells, useful nucleic acid information such as viruses, microorganisms, or circulating DNA is also lost, leading to experiment or detection failures.
The HiPure Blood DNA Kits series provided by Magen Company uses silica gel column purification technology, which can directly lyse whole blood samples without the need for white blood cell separation. Whole blood samples are directly mixed with lysates and proteases, resulting in the inactivation of pathogens, greatly reducing the infectivity, environmental pollution, and the chance of operators being infected. Due to the direct lysis and digestion of samples, except lymphocyte DNA, other circulating DNA as well as DNA from viruses and microorganisms, can also be recovered.
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 2ml blood and 200mg tissue using Midi column
Applications
PCR, southern bolt and virus detection, etc
Purification method
Midi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples
Sample amount
0.2-2 ml
Elution volume
≥300μl
Time per run
≤80 minutes
Liquid carrying volume per column
4ml
Binding yield of column
1mg
Principles
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 protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
Fast – without separation of leukocytes, organic extraction or ethanol precipitation
Simple – all nucleic acids can be obtained by direct digestion
Wide applicability – handle a variety of liquid samples
Kit Contents
Contents
D311302
D311303
Purification Times
20
100
HiPure gDNA Midi Columns
20
100
15ml Collection Tubes
40
200
Buffer ATL
50 ml
250 ml
Buffer AL
50 ml
250 ml
Buffer GW1*
22 ml
110 ml
Buffer GW2*
12 ml
50 ml
RNase A
20 mg
90 mg
Proteinase K
100 mg
440 mg
Protease Dissolve Buffer
10 ml
30 ml
Buffer AE
20 ml
120 ml
Storage and Stability
Proteinase K, RNase A should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:
