Description

Specifications

Clone	IHC269
Source	Mouse Monoclonal
Positive Control	Spleen
Dilution Range	1:200

TIGIT is an immune receptor present on some T cells and Natural Killer cells. TIGIT binds with high affinity to the poliovirus receptor (PVR) which causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Through the CD226/TIGIT-PVR pathway, TIGIT regulates T cell mediated immunity. In cancer, TIGIT and PD-1 have been shown to be over-expressed on tumor antigen-specific CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) from individuals with melanoma. Blockade of TIGIT and PD-1 led to increased cell proliferation, cytokine production, and degranulation of tumour antigen-specific CD8+ T cells and TIL CD8+ T cells. It can be considered an immune checkpoint.

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

This product speeds and simplifies nucleic acid size selection for fragment library preparation for next generation sequencing.

Details

Specifications

Features	Specifications
Main Functions	Selectively recover DNA from PCR products and enzymatic reaction solution （Replace Beckmen or agencourt SPRISelect）
Applications	Prepare DNA library for second generation sequencing
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	DNA products, restriction endonuclease systems, or other enzymatic reaction solution
Sample amount	Appropriate
Recovery	80%
Operation time	≤50 minutes

Principle

The MagSelect method contains magnetic particles in an optimized binding buffer to selectively bind DNA fragments 100bp and larger to paramagnetic beads. Excessprimers, nucleotides, salts, and enzymes can be removed using a simple washing procedure. The result is a more purified PCR product.

Advantages

• High recovery – up to 80%
• High throughput – using magnetic beads purification technology

Kit Contents

Contents	XP-5	XP-50	XP-500
MagSelect Beads	5 ml	50 ml	500 ml

Storage and Stability

MagSelect XP should be stored at 2-8°C upon arrival and is stable up to 18 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. Shake the reagent well before use. It should appear homogenous and consistent in color.

DO NOT FREEZE.

This product speeds and simplifies nucleic acid size selection for fragment library preparation for next generation sequencing.