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.
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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
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R4316 HiPure Circulating DNA/RNA Kit
Product Info
Document
Product Info
Introduction
This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection. HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.
Details
Specifications
Features
Specifications
Main Functions
Isolation both Circulating DNA/RNA (include miRNA) from 1-5 ml serum and plasma
Applications
qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method
Midi spin column
Purification technology
Silica technology, DNA filtration technology
Process method
Manual (centrifugation or vacuum)
Sample type
serum, plasma, and other cell-free liquid samples
Sample amount
1-5 ml
Elution volume
≥20μl
Time per run
≤100 minutes
Liquid carrying volume per column
4 ml
Binding yield of column
1 mg
Principle
This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.
Advantages
High yield – most optimized process to obtain maximum free RNA and small RNA
High concentration – low elution volume (>20μl) to ensure nucleic acid concentration
High purity – low alcohol combination, completely remove inhibitor and protein pollution
High recovery – silica gel column technology can recover nucleic acid molecules at the level of PG
Large volume – 1-2ml serum and plasma samples can be processed at one time
Kit Contents
Contents
R431602
D431603
Purification Times
50 Preps
250 Preps
HiPure RNA Micro Columns
50
5 x 50
HiPure Viral Midi Columns
50
5 x 50
15 ml Collection Tubes
50
5 x 50
2ml Collection Tubes
50
5 x 50
Buffer CFL
150 ml
2 x 375 ml
Buffer CFP
30 ml
150 ml
Buffer MGW1*
100 ml
2 x 250 ml
Buffer RW2*
2 x 50 ml
5 x 100 ml
RNase Free Water
10 ml
50 ml
Storage and Stability
The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Document
This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection. HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.
Programmed Death-Ligand 1 (PD-L1), also known as CD274 or B7 Homolog 1 (B7-H1), is a transmembrane protein involved in suppressing the immune system and rendering tumor cells resistant to CD8 T cell-mediated lysis through binding of the Programmed Death-1 (PD-1) receptor. Overexpression of PD-L1 may allow cancer cells to evade the actions of the host immune system. In renal cell carcinoma, upregulation of PD-L1 has been linked to increased tumor aggressiveness and risk of death, and, in ovarian cancer, higher expression of this protein has lead to significantly poorer prognosis. PD-L1 has also been linked to systemic lupus erythematosus and cutaneous melanoma. When considered in adjunct with CD8 tumor-infiltrating lymphocyte density, expression levels of PD-L1 may be a useful predictor of multiple cancer types, including stage III non-small cell lung cancer, hormone receptor negative breast cancer, and sentinel lymph node melanoma.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.
Details
Specifications
Features
Specifications
Recommended application
Plasmid Maxi preparation
Preservation conditions
Room temperature
Stability
Up to 4 years
Filter membrane
High quality glass fiber filter GF/B, 8 layers
Membrane aperture
1.0 μm
Maximum binding yield of plasmid
1 mg
Maximum yield of alcohol mediated binding
5 mg
Plasmid yield
Up to 1mg
Single liquid carrying capacity of column
20 ml
Minimum elution volume
800 μl
Withstand centrifugal force
3,000-5,000 x g
Centrifuge
Low speed centrifuge for 50ml centrifuge tubes, >3000 x g, swing-out Rotor
Adsorption Mechanism
Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silicagel membrane, while other pollutants, mainly proteins, are removed by membrane washing.
Ordering information
CAT.No.
Product Name
Package
C13123
HiPure DNA Maxi Column III (8 x GF/B)with 50ml Collection Tubes
100/Bag
Purchase Guide
Item No.
Product Name
Membrane type/number of layers
Collection tubes
Plasmid DNA binding capacity (Physical adsorption)
Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.
Document
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
