Mi Sequence

Why sequence the immune repertoire?

Structure of T cell and B cell receptors. T cell receptors are made up of two polypeptide chains that together compose one antigen binding region. B cell receptors are made up of four peptides – two light chains and two heavy chains – that comprise two antigen-binding regions.

B cell and T cell structure and function

Germline configuration of antibody gene including V(D)J segments and constant regions with zoom in on the variable region and CDR3 and diagram of fully-spliced VDJ RNA

Diversity and differentiation in the adaptive immune system

iRepertoire eBook 1: An Intro to Immunology and Immune Sequencing cover thumbnailiRepertoire eBook 1: An Intro to Immunology and Immune Sequencing. Fill out the form to download.

eBook 1: An Intro to Immunology and Immune Sequencing

Next-generation sequencing workflow: 1) Extraction 2) Library Prep 3) Sequencing 4) Analysis

NGS overview: from sample to sequencer to results

Simple graphic describing the difference between high and low coverage regions in next generation sequencing: higher coverage equates to more reads

NGS considerations: coverage, read length, multiplexing

Clonotype: A particular CDR3 sequence. Repertoire: All of the clonotypes within a particular BCR or TCR chain. Adaptome: All of the repertoires for each of the 7 different chains.

The immune repertoire and adaptome

Technology Overview

arm-PCR

dam-PCR

iRepertoire’s primer systems

MiSeq machines

Beyond template switching

iRepertoire's eBook 2: Intro to immune sequencing with iRepertoire thumbnail

eBook 2: Immune sequencing with iRepertoire

pcr strip and biological microscope

Choice of starting template: RNA vs gDNA vs cDNA

Clonotype distribution in one person. Long-tailed negative distribution with number of clonotypes on the y-axis and nuber of aliquots on the x-axis. When immune repertoire sequencing, you want to cover this highly variable CDR3 region.

How to perform immune repertoire sequencing

MiSeq machines

Choosing your read depth

DNA samples are loaded to 96-well plate for PCR analysis

Pooling samples

graphic of cancer cell, shown in orange, and lymphocytes, shown around the cancer cell in blue

Cell-sorting Guide

Blood collection and sample considerations

Sample submission guide – bulk repertoire

Biotech sample brown cardboard shipping package

Shipping samples, receiving products, and submitting data

iRepertoire eBook 3: Planning your immune repertoire sequencing experiment

eBook 3: Planning your immune repertoire sequencing experiment

Product Manuals

iR-Complete: Dual Index Primer Kit User manual for gDNA

iR-Complete

iR-Complete: Dual Index Primer Kit User Manual for RNA

iR-Complete

Contact us for iRock manuals

Guides

iPair Analyzer™ User’s Guide

Data Analysis and iPair

iRweb: Data Analysis Guide

Data Analysis and iRweb

Production Barcodes

Barcodes

Data Sheets

iR-Complete gDNA Mixed Safety Data Sheets (SDS)

iR-Complete

iR-Complete RNA Mixed Safety Data Sheets (SDS)

iR-Complete

iR-Profile Reagent Kit Mixed Safety Data Sheets (SDS)

iR-Profile