Description
SeqMatic’s TailorMix dual-indexed PhiX control library is a 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 200uL of single stranded library prediluted in HT1 buffer to a concentration of 20pM, ready for spike in.
Note: Non-denatured PhiX is available for platforms using ExAmp cluster generation (NovaSeq, HiSeq X and HiSeq 3000/4000). Denatured PhiX is not compatible with those platforms.
Features
Positive Control Library for Illumina NGS
- Compatible with most Illumina NGS systems including HiSeq 2500, MiSeq, and NextSeq
- Note: Use non-denatured PhiX for NovaSeq HiSeq X and HiSeq 3000/4000
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 dual-index sequences (can also be used in single index runs)
- Compatible with all SeqMatic and Illumina sample prep kits
Easy to use
- Supplied as a single stranded 20pM stock in HT1 buffer
- Library ready for immediate spike in
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:
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 peace-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.
Product Support
User Guide:
User Guide (PDF): Updated April 2018
Sample Sheet Setup
The TailorMix Indexed PhiX control library contains dual indexed barcodes which can be demultiplexed with all Illumina compatible demultiplexing software.
The TailorMix Dual-Indexed PhiX control library contains dual-indexed barcodes which can be demultiplexed with all Illumina compatible demultiplexing software. To monitor the percentage of PhiX reads in a sequencing run, include the TailorMix Dual-Indexed PhiX barcodes on the sequencing Sample Sheet for demultiplexing.
If the TailorMix Dual-Indexed PhiX barcodes are not included on the sequencing Sample Sheet, 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 Dual-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 MiniSeq and NextSeq: 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
The TailorMix Dual-Indexed PhiX Control Library is stable when stored at -20°C. Storage at -80°C has shown improved stability and is recommended for long term storage.
Best Practices
• Do not store the TailorMix Dual-Indexed PhiX Control Library in frost-free freezers.
• Aliquots should be made to allow for use to minimize repeated freeze/thaw cycles.
• Determine the appropriate aliquot volume based on usage.
• Aliquots that are in use can be stable for up to one month if kept on ice at all times during use and stored at temperature of -20°C or lower when not in use.
