Combine index sequences so that signal is present in at least one. Truseq nano dna libary prep reference guide 15041110 d author. Truseq dna exome formerly truseq exome kit illumina. The automated maestro truseq stranded rna cdna prep application follows the illumina protocol with a few minor modifications see figure 2. Combine the contents of each well of column 1 into well a2 of the pdp plate. Product documentation available for download in pdf from the illumina website. The second thirdgeneration technology, introduced in 2012, was the moleculo protocol which is now marketed as illumina truseq synthetic long reads19. By combining the truseq exome kit with illumina systems using sequencing by synthesis sbs technology, researchers can identify true coding variants and minimize falsepositive and falsenegative calls. Summary the truseq nano dna sample preparation kit optimizes the truseq workflow to deliver a lowinput sample preparation method for any sequencing application.
Infinium methylationepic kit methylation profiling array. A fundamental feature of all library preparation methods for ngs is their speed, with decreasing laboratory and machine run time. The collibri ps dna library prep kit for illumina systems detects snps from 1 ng input amplified with similar sensitivity to pcrfree protocols. Automating the illumina truseq stranded mrna library prep. Truseq stranded total rna with illumina ribozero plus. Instructions for preparing libraries using a truseq nano dna library prep kit. Each truseq dna pcr free lt library prep kit contains adapter index tubes. Microsoft word understanding illumina truseq adapters. Truseq dna and rna library prep kits include gelfree protocols that eliminate the timeintensive gel purification step found in other methods, making the process more consistent and fully automatable. Truseq rna sample preparation v2 guide 15026495 f illumina. Truseq dna nano interrogate low input samples illumina.
Adjust the concentration of the chip enriched dna 10ng50ul in nuclease free water. Truseq nano dna library prep checklist documentation. Thenumberofsamplesthatcanbepooledperrundependsonthescopeandgoalofagivensequencingproject. Using this approach, 10kbp molecules of dna are clonally amplified and barcoded before sequencing with a short read instrument, so that long. This means combining one indexed oligo with the universal oligo, and. Automated illumina truseq nano dna sample preparation on.
Illumina will not officially support any method other than its own, and we strongly advise using their method, or at the very least, make only minor adjustments. This document and its contents are proprietary to illumina, inc. Tableofcontents chapter1overview 1 introduction 1 dnainputrecommendations 1 additionalresources 2 chapter2protocol 3 introduction 3 tipsandtechniques 3. Automating the illumina truseq stranded mrna library prep protocol with the tecan freedom evo ngs workstation author. Illumina nextera xt sample preparation protocol chmi. Truseq stranded mrna sample preparation guide 15031047 e. Truseq rna exome formerly truseq rna access library prep kit. The goal of this protocol is to add adapter sequences onto the ends of dna fragments to generate multiplexed single read or paired end sequencing libraries. Product documentation in pdf is available for download from the illumina website. Truseq nano dna libary prep reference guide 15041110 d. Wholeexome enrichment with the illumina truseq exome. Users guide illumina truseq rna library preparation on the sciclone ngs liquid handler compatible with.
The library preparation protocol requires one kit from each component, depending on sample requirements. Truseq enrichment guide ut southwestern medical center. For 711plex pools, combine any of the 26plex pools. This process also describes how to combine multiple libraries with different. The gelfree protocol for truseq dna library preparation is available. This reduced input requirement extends the accessibility of truseq data quality to an even broader range of applications and study designs figure 2. Set up the indexes you will be using and arrange them in the truseq index plate fixture see manual for images.
You do not want the ampure beads to appear cracked or crusty. Thirdgeneration sequencing and the future of genomics. Automated illumina truseq stranded total rna library. The illumina truseq small rna sample preparation protocol is used to prepare a. Quality control in illumina sequencing workflows using the tapestation system. Shear dna to a size distribution peaking around 400600 bp using a diagenode bioruptor or covaris. Product documentationavailable for download in pdf from the illumina website. Combine sample purification beads and pcr grade water in a tube to create a diluted. The goal of this protocol is to enrich for sequences in a solution using two rounds of hybridizations. With the discontinuation of the market leader for library preparation methods, illumina s truseq dna v2 kit, a gap has been created for a new method to become the most widely used kit.
If you are using custom truseq style sequencing adapters i. Use 1ug of total rna to initiate protocol quantity may vary by sample type reference protocol. Lab efficiency comparison manual neoprep reagent kit truseq stranded mrna library prep kit truseq. Truseq nano dna library prep kit for neoprep author. Lt protocol 1 performed using the truseq small rna sample prep kit illumina sets. This protocol explains how to convert total rna into a library of template molecules of. Truseq stranded total rna with illumina ribozero plus rrna depletion reference guide documentation. Truseq and other illumina library prep technologies ensure sample accuracy. For each sample, combine the following volumes in a pcr tube. Allow the tubes containing the ampure beads to dry for approximately 5 minutes or until there is no longer a smell of etoh. Truseq nano dna libary prep protocol guide 15075697 a. Illumina subject gain a clear and comprehensive view of the transcriptome with a streamlined, costefficient, and scalable solution for mrna or wholetranscriptome analyses. Table 2 kit, sample number, and protocol recommendations.
This protocol is based on the illumina truseq exome enrichment platform, which captures 62 mb of the human exonic regions using 95base dna probes. This provides a lowcost solution for analyzing human rna isolated from formalinfixed, paraffinembedded ffpe tissues and other lowquality samples. This application note describes an automated illumina qualified protocol developed for parallel processing of up to 48 samples using truseq stranded mrna kits with the tecan freedom evo ngs workstation. Each sequencing workflow step is optimized to deliver accurate, highquality data. Illumina truseq adapters, bfidt adapters, itrue adapters see. Tableofcontents chapter1overview 1 introduction 1 process 1 rnainputrecommendations 4 additionalresources 6 chapter2protocol 7 introduction 7 tipsandtechniques 7. Automated illumina truseq rapid exome library construction. In my experience, it takes about 7 minutes for tubes to airdry in a low humidity environment. In addition, two nontemplate control samples ntc were prepared.
F march 2014 truseq rna sample preparation v2 guide. This chapter describes the truseq dna methylation kit protocol. The illumina free adapter blocking reagent is an optional reagent that can be used to treat most types of sequencing libraries to reduce index hopping levels. Illumina remains committed to providing you with highquality support and service. Truseq stranded mrna library prep kit for neoprep illumina. Multiple platforms are available for wholeexome enrichment and sequencing wes. Preparation of sequencing libraries from dna or cdna for sequencing on an illumina platform. Combine the set a and set b indexes to pool up to 24 libraries. The proven truseq nano dna biochemistry is now supported by the neoprep library prep system, an integrated solution for preparing libraries for sequencing on any illumina sequencing platform, making library prep far easier than ever before.
The onetube chemistry and protocol improves the efficiency and consistency of library construction, and yields libraries of similar or better quality than those produced with the kapa hyperprep kit from covarissheared dna. Dna fragmentation and library construction protocol for the rapid preparation of libraries for illumina sequencing. C ollibri ngs library preparation kits for highthroughput. Truseq nano dna library prep usersupplied consumables and equipment documentation. This protocol incorporates the withbead ampure cleanup protocol, initially. Euc and ltf merged into a single document per protocol.
The infinium methylationepic kit is the recommended replacement product for the humanmethylation450 beadchip kit, which has been discontinued. Clarified pdp plate type options for ls protocol are a 0. Reagents provided in the truseq exome enrichment or truseq custom enrichment kits subsequently prepare the library for sequencing targeted regions on the illumina sequencing platform. Truseq nano dna libary prep protocol guide 15075697 a author. Illumina uses onetrust, a privacy management software tool, to handle your request. Reagents provided in the truseq exome enrichment kit subsequently prepare the library for sequencing targeted. Truseq dna pcrfree library prep kits provide uniform coverage for. Input recommendations for tmb, msi, and small variant analysis with trusight oncology 500. Illumina experiment manager user guide documentation. Custom protocol selector generates customized, endtoend instructions.
Added truseq stranded total rna ht sample prep kit content. Table 17 truseq stranded mrna lt sample prep kit set a indexed adapter. Note that sequencing nextera xt libraries on certain instruments the hiseq 2000, hiseq 15002500 in. In this technical note, we present the automation of the stranded mrna sample preparation kit protocol on biomek i7 dual hybrid multichannel 96, span8 genomics workstation. Truseq nano dna library prep kit for neoprep illumina. Additional product documentation in pdf is available for download from the illumina.
Sequencingready libraries were prepared from depleted rna using the truseq stranded total rna library prep gold kit illumina, cat no. Illumina, truseq, the pumpkin orange color, and the streaming bases design are. Combine the following in a separate, sterile, nucleasefree, 200 l pcr tube. All control samples were processed using the full illumina truseq. Quality control in illumina sequencing workflows using the. Truseq rna exome, previously known as the truseq rna access library prep kit, converts total rna into template molecules of known strand origin, followed by sequencespecific capture of coding rna. Truseq rna and dna sample preparation kits v2 illumina. Library preparation illumina library prep protocol 2. Coriell na12878 dna was converted into a sequencing library by the collibri ps or pcrfree ps library prep kit and sequenced on a 2 x 150 bp paired end run on an illumina novaseq 6000 system. Zero gold rrna depletion protocol to remove human cytoplasmic and mitochondrial rrna illumina, 48 samples, cat no. You may receive emails through the onetrust system as your request is processed.
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