Purify sequencing extension products from dye terminators, primers and other contaminants
Also purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations.
High recovery
Fast and efficient spin column format
Also available in 96 well format for high throughput
This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.
The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up. Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
Component
Cat. 34500 (50 preps)
Cat. 34400 (2 x 96-well plate)
Binding Buffer C
30 mL
3 x 30 mL
Wash Solution A
12 mL
2 x 20 mL
Elution Buffer B
8 mL
2 x 15 mL
Spin Columns
50
–
Collection Tubes
50
–
Elution Tubes (1.7 mL)
50
–
96-Well Plate
–
2
Adhesive Tape
–
4
96-Well Collection Plate
–
2
96-Well Elution Plate
–
2
Product Insert
1
1
Other Products
D3018 HiPure Universal DNA Kit
Product Info
Document
Product Info
Introduction
This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from tissue / blood / body fluid / swab /dry blood spots
Applications
PCR, qPCR, southern bolt and virus detection, etc
Purification method
Mini 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
Solid tissue: 1-10mg; Anticoagulant blood: 200μl
Elution volume
≥20μl
Time per run
30 – 60 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
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
D301802
D301803
Purification Times
50
250
HiPure DNA Mini Columns I
50
250
2ml Collection Tubes
100
500
Buffer ATL
30 ml
150 ml
Buffer AL
30 ml
150 ml
Buffer GW1
22 ml
88 ml
Buffer GW2
12 ml
50 ml
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
1.8 ml
10 ml
Buffer AE
15 ml
60 ml
Storage and Stability
Proteinase K 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. 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.
Experiment Data
Document
This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.
This is a high-quality (commercial grade) version of the T4 DNA Ligase. T4 DNA Ligase is recombinantly produced in E. coli. ArcticZymes’ T4 DNA Ligase is extensively tested for contaminating DNase and RNase activities as well as residual host-cell gDNA.
Key Features
ATP and Mg2+ dependent dsDNA ligase
Easily heat-inactivated at 70°C for 10 minutes
Extensively tested for contaminating DNase and RNase activities as well as residual host-cell gDNA
Applications
Ligation of dsDNA
NGS library prep
Molecular cloning
Figures
Document
T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.
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 whole blood, tissue and culture cells.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 10ml blood and 1g tissue using Maxi column
Applications
PCR, southern bolt and virus detection, etc
Purification method
Maxi 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
3-10ml
Elution volume
≥700μl
Time per run
≤90 minutes
Liquid carrying volume per column
4ml
Binding yield of column
5mg
Principle
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
Pertinence – specially designed for isolating DNA from 3-10ml blood and related body fluids
Wide applicability – handle a variety of liquid samples
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.
*Note：Leukocyte protocol can be used when large volume whole blood samples need to be processed. Whole blood was treated with red blood cell lysate, and white blood cells were obtained by centrifugation before extraction
Document
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:
