The RNA Seq Library Prep Kit was developed for construction of high quality NGS  libraries for next generation sequencing (illumina and MGI Platforms). RNA Sequencing is a very powerful tool to analyze transcriptome such as gene expression and transcription regulation, splicing characterization, mutation and variation detection etc. The kit needs purified RNA (example: rRNA depleted RNA or polyA mRNA) as input for library construction. Library multiplexing is possible with different types of indexes.

RNA Seq Library Prep Kit Workflow

Three index types are available for the illumina platform kits:

Non-index (illumina Cat.# 30055): Libraries do not have index.

Index (illumina Cat.# 30056): A unique barcode sequence with 6 bases has been included in each of the index primers. RNA Sequencing library multiplexing is possible with up to 48 samples. Index information can be downloaded here.

Unique dual index (illumina Cat.# 30057): RNA Sequencing library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a 4-Base Difference Index System. The system can generate indexes with at least 4 bases different from others in the 8-base indexing region. the unique dual indexing primers identify sequencing errors such as index hopping, mis-assignment, and de-multiplexing errors. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34112).

Features

• • • Quick protocol
      • Libraries will be ready in 2 hours
      • Hands-on time is only ~10 minutes
    • Guaranteed quality
      • High yield
      • Uniform coverage of transcripts
    • Simple workflow
    • Input purified RNA amount: From 3 ng to 100 ng

Comparison of the protocol time: BioDynami RNA Seq Library Prep Kit vs other vendors’ kits. Hands-on time and walk-away time were indicated.

Library size and distribution of BioDynami kit, 20 ng and 3 ng of polyA mRNA as input.

Comparison of library yield and duplication rate under the same condition: 20 ng and 3 ng of polyA mRNA were used as input. PCR cycle numbers were indicated.

Comparison of coverage of transcripts: 20 ng of polyA mRNA were used as input. BioDynami kit has more uniform coverage

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 whole blood, tissue and culture 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.
• 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
• Economy – excellent cost effectiveness performance

Kit Contents

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.

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

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, urine, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.

Details

Specifications

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA / RNA is released into the lysate. Transfer to an adsorption plate and filter column. DNA/RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA / RNA was finally eluted with low-salt buffer (10 Mm Tris, pH 8.0).

Advantages

• Fast – several samples can be extracted in 20 minutes by column method
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG
• High yield – carrier RNA contained in the product maximize the recovery of trace nucleic acid

Kit Contents

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (Proteinase K up to 8 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 18 months under these conditions.

Experiment Data

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, urine, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.