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 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 [email protected] TailorMix Ready to Use Indexed PhiX Control July 2014 Contents TailorMix Ready to Use Indexed PhiX Control 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 [email protected] TailorMix Ready to Use Indexed PhiX Control July 2014 Procedures: 1. 2. 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: [email protected] Enabling Accurate NGS www.SeqMatic.com
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