TailorMix Dual Indexed PhiX Control Library (Nondenatured)

$150.00

Barcoded PhiX control library for high quality NGS without PhiX contamination.

Each tube contains 10uL at 10nM.

SKU: TM-502-ND Category:

Description

SeqMatic’s TailorMix dual 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 runs, enabling the generation of cleaner raw data and the option to analyze the PhiX reads for quality control purposes. Each tube contains 10uL of double stranded library at a concentration of 10nM and can be used as a direct replacement for Illumina PhiX V3.

Features

Positive Control Library for Illumina NGS

  • Compatible with all Illumina NGS systems including HiSeq 4000, HiSeq 2500, MiSeq, and NextSeq
  • Can be used in place PhiX V3

High Flexibility

  • Positive control library used to monitor instrument error rates and read quality
  • High diversity library can be used to supplement low nucleotide diversity sequencing runs

Unique dual index barcodes

  • Custom designed index sequence
  • Compatible with all SeqMatic and Illumina sample prep kits

Easy to use

  • Supplied as a double stranded 10nM stock
  • Direct replacement for Illumina PhiX V3

Recommended For:

  • 16S amplicon sequencing
  • Single PCR amplicon sequencing
  • Viral genome sequencing
  • Low diversity libraries
  • Highly multiplexed pools
  • Single sample runs
  • Unindexed libraries

 

Sample Data:

PhiX Data

Figure 1: Percentage of sequencing reads mapping to the PhiX 174 genome using Indexed PhiX compared against Illumina PhiX V3.

 

Customer Testimonials:

“PhiX contamination in our data used to be a significant issue given that we were sequencing viral DNA extracted from patient biopsies.  We no longer see PhiX contamination with your product, which ultimately streamlines our data processing and gives us additional piece-of-mind regarding the validity of our data.”

Eric Sherman
University of Pennsylvania

 

“My lab was noticing large amounts of bleed-through of Illumina’s PhiX v3 into our indexed samples. Seqmatic’s Indexed PhiX dramatically reduces this bleed-through and allows us to estimate our incidence of sequencing crossover contamination.”

Customer C.F.
San Francisco, CA

 

Citations & References:

Shechner, David M., et al. “Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display.Nature methods 12.7 (2015): 664-670.

Documentation

User Guide:

User Guide (PDF): Updated April 2017

Sample Sheet Setup

The TailorMix Indexed PhiX control library contains dual indexed barcodes which can be demultiplexed with all Illumina compatible demultiplexing software.

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. Simply add a new line to your sample sheet file containing the sequences below as your index barcodes. For single index read sequencing runs, only use the i7 Index 1 barcode sequence. The sequence length may also be adjusted as necessary to match the length of the index read.

i7 Index 1 barcode: TCGAATGA
i5 Index 2 barcode for MiSeq and HiSeq 2000/2500: GACATGCG
i5 Index 2 barcode for NextSeq and HiSeq 3000/4000: CGCATGTC
 

Input Recommendations

Note: Optimal ratio of Dual-Indexed PhiX control library in the sequencing run could be different for each amplicon library.

Library Type Input Recommendation
  • TruSeq DNA or RNA
  • Nextera or ScriptSeq
  • TailorMix DNA or RNA
1% of final loading concentration
  • TruSeq Small RNA
  • TailorMix miRNA
5-10% of final loading concentration
  • PCR Amplicons (e.g. 16S, ITS, etc.)
15-25% of final loading concentration
  • Control Lane
  • PhiX Only Run
100% of final loading concentration

 

Storage Recommendations

An unopened tube of Indexed PhiX should be stable at -20°C for six months. If you plan on using the product for many freeze/thaw cycles, we advise aliquotting the PhiX into smaller tubes and storing them at -20°C. Tubes that are in use can be stable for up to one month if kept on ice at all times during use and stored at -20°C when not in use.

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