The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.

NGS Single Stranded DNA Library Prep Kit workflow

Three index types are available for the kit:

Non-index (Cat.# 30081): Libraries do not have index.

Index(Cat.# 30082): Each of the index primers has a unique 6-base index sequence that can be used to identify libraries. ssDNA library multiplexing is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30083): ssDNA sample multiplexing up to 96 libraries is possible with unique dual indexes. We have developed a Four-Base Difference Index System. This makes it possible to generate indexes with at least 4 bases different from each other in the 8 bases index length. The unique dual indexing primers remove NGS errors (example: de-multiplexing errors, read mis-assignment, index hopping etc). Index information can be downloaded here.

Kit advantages:

• • • Quick and simple Protocol
      • Quick: Total protocol time is only 1.5 hours
      • Simple: Hands-on time only 10 minutes
    • Easy procedure based on:
      • The ready-to-use master mix: Made reaction setup easy
      • Less reaction components: Simplify the reaction preparation
    • Less magnetic beads needed: Reduced more than 50% of the beads for cleanup steps
    • Directional library
    • Single stranded DNA as input: From 50 ng to 500 ng

NGS Single Stranded DNA Library Prep Kit has similar library conversion efficiency and yield as compared to a regular DNA library prep kit.

Alignment rate and duplication rate: comparison of single stranded DNA library kit versus double stranded DNA library prep kit.

The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.

Introduction

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.

At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.

This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Details

Specifications

Principle

Stool sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. Humic acid, proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Advantages

• High purity – unique adsorbent can completely remove inhibitory factors
• High concentration – maximum extraction of total DNA from stool samples
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Storage and Stability

Proteinase K and Buffer PCI 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.

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.

At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.

This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Introduction

This product is suitable for rapid extraction of 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

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High purity – OD 260 / 280 : 1.7- 1.9, OD 260 / 230 : 1.5 – 2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• High yield – most optimal process, ensuring the recovery up to 90%
• Strong processing ability – samples including animal blood, cultured cells, animal tissues, etc.

Kit Contents

Storage and Stability

Proteinase K, RNase A, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 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.

Experiment Data

This product is suitable for rapid extraction of 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.