The Multiplexing Index Primers contain primer mix for multiplexing library samples for Next Generation Sequencing (NGS) on the illumina platform. Multiplexing of NGS library samples will reduce sequencing costs by pooling multiple NGS libraries into a single flow cell lane.

With library multiplexing, unique index sequence is added to individual sample during NGS library preparation.  Therefore, each DNA molecule can be identified after pooling of multiple samples based on the index information they have.

Each of our index primers contains a unique index sequence with 6 bases that can be used to identify libraries. Library multiplexing up to 48 samples is possible.

Multiplexing Index Primers (illumina platform): Even distribution of 48 samples using index primers. 48 libraries were made using the BioDynami NGS DNA Library Prep Kit (Cat. # 30009) and the BioDynami Multiplexing Index Primers (Cat. # 30072). Libraries were pooled at equal concentration and sequenced on the illumina HiSeq 2500. The numbers of reads from 48 libraries were analyzed.

List of index sequence for the primers (each of the index primer mix contains universal primer and one of the index primers). Index number and the index sequence are listed.

List of indexes can be downloaded Here.

Sequence of the final library with index location:

The Multiplexing Index Primers contain primer mix for multiplexing library samples for Next Generation Sequencing (NGS) on the illumina platform. Multiplexing of NGS library samples will reduce sequencing costs by pooling multiple NGS libraries into a single flow cell lane.

Introduction

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, whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Details

Specifications

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.
• High throughput – 96 samples can be processed simultaneously

Kit Contents

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.

Purchase Guide

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:

Introduction

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.

Details

Specifications

Principle

Highsalt mediated binding: in the solution containing 2-4M guanidine isothiocyanate, Magpure particles can selectively recover DNA molecules, and impurities such as protein polysaccharides are not adsorbed.

Alcohol mediated binding: in the solution containing guanidine salt and alcohol (>25%), Magpure particles can selectively recover DNA/RNA molecules, and proteins and other impurities are not adsorbed.

After biological samples are treated with digestive solution or lysis Buffer, DNA/RNA is released from cells, organelles and protein complexes (ribosomes and nucleosomes) into reagents. After Magpure particles and binding solution are added, DNA/RNA is adsorbed to the surface of Magpure particles to form DNA/RNA bead complex. Under the action of the magnetic field, the magnetic beads are separated and collected, and the impurities such as protein are removed with the waste liquid. After two or three steps of further cleaning, the DNA/RNA magnetic bead complex is resuspended in sterilized water or TE buffer, and the DNA/RNA falls off from the surface of the magnetic beads, so as to achieve the purpose of purification.

Ordering information

Purchase Guide

• The MagPure magnetic-particle technology combines the speed and efficiency of silica-based DNA purification with the convenient handling of magnetic particles. DNA binds to the silica surface of the magnetic particles in the presence of a chaotropic salt. DNA bound to the particles is then efficiently washed, considerably improving the purity of DNA. High-quality DNA is eluted. The automated purification procedure completely removes enzymes, nucleotides, and other contaminants and inhibitors. Purified DNA is suitable for direct use in downstream applications, such as sequencing and microarray analysis.

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.