Indexed PhiX Control Library Introduction

Indexed PhiX Control Library
Catalog Number: TM-501
Introduction
SeqMatic’s indexed PhiX control library is a prepared
adapter ligated library used as a control for Illumina sequencing
instruments. Our custom indexed library eliminates PhiX
contamination in multiplexed sequencing run.
Features
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100% Compatibility: Can be used in place of Illumina PhiX V3
in all Illumina systems.
High diversity: Library contains a randomized insert library
with 45% GC content and can be used to supplement low
diversity sequencing runs.
Unique index barcode: Custom designed index sequence is
compatible with all Illumina sample prep kits.
Easy to use: Supplied as a ready to use 20pM single stranded
library for immediate spike in.
29528 Union City Blvd.
Union City, CA 94587
Tel: 510.870.0965
www.seqmatic.com
info@seqmatic.com
TailorMix Ready to Use Indexed PhiX Control July 2014
Contents
TailorMix Ready to Use Indexed PhiX Control
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Sample type: Single stranded Illumina adapter ligated
library
Concentration: 20pM
Volume: 200µL
Insert size: 380bp
I7 Index barcode: TCGAATGA
I5 Index barcode: TCTTTCCC
Consumables Preparation
Please make sure all consumables and equipment are available before
starting this experiment.
Consumables and Equipment
Micropipettor
Supplier
General lab supplier
Micropipettor Tips
HT1 Buffer
General lab supplier
Illumina
Microcentrifuge tubes
General lab supplier
29528 Union City Blvd.
Union City, CA 94587
Tel: 510.870.0965
www.seqmatic.com
info@seqmatic.com
TailorMix Ready to Use Indexed PhiX Control July 2014
Procedures:
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3.
Denature and dilute your sequencing libraries according to Illumina’
standard operating procedures.
Mix the 20pM Indexed PhiX control with your denatured and diluted
sequencing library.
a. For standard TruSeq and Nextera libraries:
i. Add 1% of the denatured Indexed PhiX control library to your
sample as a positive control for QC purposes.
b. For miRNA and Small RNA libraries:
i. Add 5-10% of the denatured Indexed PhiX control library to
your sample as a positive control for QC purposes.
c. For low diversity amplicon libraries:
i. On MiSeq: Add 20% of the denatured Indexed PhiX control
library to your sample as a positive control for QC purposes
and for increasing diversity of your samples.
ii. On Genome Analyzer, and HiSeq: Add 40% of the denatured
Indexed PhiX control library to your sample as a positive
control for QC purposes and for increasing diversity of your
samples.
d. For dedicated control lanes on Genome Analyzer and HiSeq:
i. Load the proper concentration of denatured Indexed PhiX
library according to Illumina’s standard operating procedures
for your instrument.
Load your sample onto the sequencing or clustering instrument
according to Illumina’s standard operating procedures for your
instrument.
Enabling Accurate NGS
www.SeqMatic.com
TailorMix Ready to Use Indexed PhiX Control July 2014
Sample Sheet Setup
Sample sheet modifications are not necessary for most
applications. The PhiX reads will be parsed into a folder named Unknown
or Undetermined along with any other reads with index sequences not
defined in your sample sheet.
For individual downstream analysis of the Indexed PhiX library,
your sample sheet can be modified to parse the PhiX reads into its own
individual set of files.
Ordering Information
Catalog #
TM-500
TM-501
Product
10nM Index PhiX Control
Library
Ready to use 20pM single
stranded Indexed PhiX
Control Library
Visit our SeqMatic.com to browse our entire line of NGS products
and services.
SeqMatic LLC
44846 Osgood Rd.
Fremont, CA 94539
www.seqmatic.com
Tel: 1.510.870.0965
Email: info@seqmatic.com
Enabling Accurate NGS
www.SeqMatic.com