*PhiX can be further adjusted based on experimentation. DATE POSTED. Target regions can be as small as 1 bp, but because designs must include 12 amplicons, you would need 12 sets of 1 bp regions. NovaSeq Series Custom Primers Guide (1000000022266 v03) ... and providing the highest level of quality, we strive to meet this challenge. FAQ: Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext ® Multiplex Oligos for Enzymatic Methyl-seq on Illumina ® sequencing instruments? The VP10 Custom Read 1 primer is required for all sequencers, while the VP14 Custom Index 2 primer is required for Optimal loading concentrations for Illumina DNA PCR-Free libraries Load Illumina DNA PCR-Free libraries onto compatible sequencing systems at optimal FILE NAME. It covers the poly(T) stretch and replaces the Multiplex Read 1 Sequencing primer. All libraries prepared with the current Illumina library preparation kits are compatible with all Illumina sequencing platforms. V1 = solve for the volume of the custom primer to be spiked in C2 = the recommended custom primer final concentration from the chart below V2 = total volume of Illumina primer in the charts below Example for MiSeq platform: 100 M * V1 = 0.5 M * 680 L V1 = 3.4 µl Important Note: The guidelines below are based on the current primer volumes. We developed a custom indexing primer that allows multiplexing of samples on the X system, including the use of a custom Libraries made using NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 2) are fully compatible with Illumina sequencing protocols for TruSeq ® HT libraries. No. The SureCell Sequencing Primer is used for Read 1 and is added to the custom primer position (consult the custom primer documentation for your sequencing instrument). AmpliSeq for Illumina custom panels range from 12 amplicons to 3,072 amplicons per pool. You may also want to spike in your custom primers. No. Here a Custom Sequencing Primer (CSP Version 2, included in the kit) is required to achieve cluster calling on Illumina machines. Files. Files. This kit eliminates the need for PhiX spike … Illumina recommends starting with higher spike-in percentages and reducing based on run performance. Type & Size. Yes, the SureCell WTA 3’ Library Prep Kit comes with the SureCell Sequencing Primer at 50 µM (in Box 3 of 4) that is compatible with only SureCell libraries and PhiX. 015 (QuantSeq 3‘ mRNA-Seq Library Prep Kit for Illumina (FWD)) 016 (QuantSeq 3‘ mRNA-Seq Library Prep Kit for Illumina (REV) with Custom Sequencing Primer) 020 (PCR Add-on Kit for Illumina) 022 (Purification Module with Magnetic Beads) 025 (SIRVs Spike-in RNA Variant Control Mixes) 026 (QuantSeq-Flex First Strand Synthesis Module) FILE INFO. Frequently Asked Questions: Fungal ITS: Illumina has published a second version of this protocol, modified to sequence and study fungal ITS sequences. The Illumina DNA PCR-Free workflow requires the use of custom sequencing primers. Qiagen offers a commercial amplicon prep kit for multiple 16S regions and ITS for which they have perfected the diversity spacer approach described above. FAQ: Do I need to spike in custom sequencing primers when sequencing libraries are made with NEBNext ® Multiplex Oligos for Illumina ® (Dual Index Primers Set 2) on Illumina sequencing instruments? 016) the Read 1 linker sequence is located at the 5’ end of the oligodT primer. HiSeq System Custom Primers Guide (15061846 v03) ... and providing the highest level of quality, we strive to meet this challenge. For QuantSeq REV (Cat. Libraries made using NEBNext Multiplex Oligos for EM-seq (96 Unique Dual Index Primer Pairs) are fully compatible with Illumina sequencing protocols for TruSeq® HT libraries. Instructions for sequencing with custom primers on the NovaSeq Sequencing System. No. Date. For more information, see the bulletin Spiking custom primers into the Illumina sequencing primers. A (G+C)-rich spike-in such as from K. radiotolerans in the setting of the new HiSeq X software is much more robust to such variation, with sequencing base quality maintained even at 2-3% spike-in levels. Name. Instructions for sequencing with custom primers on any HiSeq system (except the HiSeq X System). Email Technical Support at techsupport@illumina.com with any questions about your particular library and platform. All orders have a minimum price equivalent to … 1000000022266 v03 )... and providing the highest level of quality, we strive to illumina custom primer spike in this challenge kit is. 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