Use the TruSeq Genotype Ne kit for targeted genotyping by sequencing for parentage, purity testing, and breeding decisions for any plant or animal species. The kit produces high-quality results for up to 5000 SNPs and medium to high sample volume (1000–100,000).
TruSeq Genotype Ne supports amplicon sizes of 150 bp and 175 bp.
Illumina recommends using 50 ng high-quality genomic DNA (gDNA). Use a fluorescence-based method, such as Qubit dsDNA Assay Kit or PicoGreen to quantify DNA. For more information on quantifying and diluting input DNA, see the TruSeq Genotype Ne Reference Guide (document # 1000000033138).
Yes. The number of PCR cycles performed depends on amplicon plexity. Using the recommended number of PCR cycles ensures sufficient yield for bead-based normalization while avoiding overamplification. For recommendations, see the TruSeq Genotype Ne Reference Guide (document # 1000000033138).
The TruSeq Genotype Ne Kit supports up to 5000 amplicons per reaction. The number of amplicons in the multiplex reaction varies depending on the experiment design, and can range from 116–5000.
The protocol requires two different magnetic stands: one for pre-PCR procedures and one for post-PCR procedures.
The recommended loading concentration depends on the sequencing system. For recommendations and instructions, see the denature and dilute libraries guide or system guide for your instrument.
The TruSeq Genotype Ne kit is customizable, so the number of samples and the sequencing system are specified when the kit is ordered. Samples are prepared in multiples of 96, and the kit can support sequencing up to 384 multiplexed samples. The actual number of libraries that can be pooled together per sequencing run depends on the number of amplicons and the desired sequencing coverage.
Illumina recommends run parameters of 1 × 121 cycles with dual-indexing. The MiniSeq, MiSeq, NextSeq 500, and NextSeq 550 systems are compatible with TruSeq Genotype Ne libraries.
Use the TruSeq Amplicon BaseSpace App or the Local Run Manager Amplicon DS analysis module to analyze TruSeq Gentoype Ne libraries.