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Cat. No.: DLQP-20 (for 20rxn)

Cat. No.: DLQP-100 (for 100rxn)

Description

SBS-NGS™ DNA Library Quick-Prep Kit (For Illumina) is a rapid library preparation kit developed specifically for the Illumina high-throughput sequencing platform. Utilizing in vitro transposition technology, the kit enables simultaneous DNA fragmentation and adapter addition in just 10 minutes with as little as 1–100 ng of DNA. This eliminates the need for the traditional, multi-step processes of fragmentation, end repair, and adapter ligation. Following this step, PCR amplification with the kit produces sequencing-ready libraries compatible with Illumina platforms. Key advantages include low DNA input requirements, simplified workflow, and significantly reduced library preparation time. It is suitable for genomic DNA, PCR amplicons, plasmids, and similar DNA samples for fragmentation, adapter ligation, and amplification.

Next Generation Sequencing (NGS)—also known as high-throughput sequencing or deep sequencing—is a new generation of DNA sequencing technology developed from PCR and microarray platforms. The experimental workflow of NGS typically includes: sample preparation, cluster generation, sequencing, and data analysis. In NGS, individual DNA molecules must be amplified into clusters of identical DNA fragments. These clusters are then synchronously replicated to enhance fluorescence signal intensity, enabling accurate sequence reading. However, as read length increases, cluster synchronization decreases, leading to reduced sequencing quality. Thus, the effective read length of NGS is limited; in paired-end sequencing, read lengths generally do not exceed 500–600 bp.

With the Tn5 Transposome, genomic DNA and other DNA samples undergo a one-step tagmentation reaction (DNA fragmentation and tagging), simultaneously fragmenting DNA and adding adapters. The resulting tagged DNA is then subjected to PCR amplification, during which gaps are filled and the library is amplified using dual-indexed PCR primers. This process generates DNA libraries compatible with Illumina NGS platforms. Subsequent purification steps, such as magnetic bead-based size selection, remove large DNA fragments, primers, and PCR reagents, yielding DNA libraries with the desired fragment size distribution for high-quality sequencing.

This kit provides a specially optimized SBS-NGS™ PCR Master Mix (2X) for library amplification. It employs a high-fidelity, fast-amplifying DNA polymerase together with an optimized reaction buffer. Users only need to add the DNA fragments with adapters obtained from the previous step along with appropriate primers. This greatly simplifies the library amplification workflow, reduces the risk of contamination during PCR operations, and ensures extremely low error rates during amplification. As a result, the library amplification process achieves high fidelity, efficiency, uniformity, and reproducibility.

The kit does not include the Dual-Indexed PCR Primers required for amplification. We provide SBS-NGS™ Dual-Index Primers of Tagmentation (For Illumina), which consist of 4 sets. Each set contains 4 types of 8-base (nt) i5-tagged P5 primers (SBS-NGS™ Nex i5 Primers) and 6 types of 10-base (nt) i7-tagged P7 primers (SBS-NGS™ Nex i7 Primers). When used individually, each set allows cross-combination of P5 and P7 primers to generate 24 unique i5/i7 index pairs. When all 4 sets are used together, up to 384 unique i5/i7 index pairs can be generated, giving users flexibility to select according to their experimental needs.

Components

• SBS-NGS™ Tn5 Transposome (20μM)
• Reaction Buffer (5X)
• Stop Buffer (5X)
• SBS-NGS™ PCR Master Mix (2X)

Application

This kit is designed for preparing DNA samples into sequencing-ready libraries specifically for the Illumina high-throughput sequencing platform. For representative results of DNA libraries prepared using our SBS-NGS™ DNA Library Quick-Prep Kit (For Illumina), please refer to the figure below.

The SBS-NGS™ Tn5 Transposome included in this kit contains the following ME and adapter sequences:

• ME: 5'-p-CTGTCTCTTATACACATCT-3'-OH

• Primer 1: 5'-OH-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3'-OH

• Primer 2: 5'-OH-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3'-OH

Note: The above adapter sequences are identical to those used in the Nextera® DNA Sample Preparation Kit (Illumina, FC-121-1030/1031) and the Nextera XT DNA Library Preparation Kit (Illumina, FC-131-1024/1096).

According to the instructions, if each reaction uses 1 μl of SBS-NGS™ Tn5 Transposome (20 μM), the small package and medium package of this kit can perform approximately 20 reactions and 100 reactions, respectively.

Storage

Store at –20 °C. Stable for one year.

Notes

• The SBS-NGS™ Tn5 Transposome (20 μM) storage solution contains 50% glycerol. It will not freeze at –20 °C. Do not store at –80 °C, as the solution will freeze; repeated freeze–thaw cycles may reduce Tn5 transposase activity.
• The SBS-NGS™ Tn5 Transposome (20 μM) is viscous. Ensure accurate pipetting, and after addition, mix thoroughly by pipetting up and down. Avoid introducing bubbles.
• The substrate of SBS-NGS™ Tn5 Transposome (20 μM) is DNA; it is not suitable for RNA experiments.
• The SBS-NGS™ PCR Master Mix (2X) must be completely thawed before use. Mix gently by inverting several times, avoiding bubble formation.
• This product is intended for research use only by trained professionals. It must not be used for clinical diagnosis or therapy, food, or pharmaceuticals, and should not be stored in residential settings.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.