Compare kapa hyper prep kit illumina, with kapa library amplification primer mix 10x from roche on. Kapa library quantification kit illumina platforms roche. Kapa sybr fast qpcr master mix 2x kit sigmaaldrich. We used the kapa stranded mrnaseq kit trad kapa to prepare libraries using the whole transcript method, and the lexogen quantseq 3mrnaseq library prep kitfwd 3lexo to prepare 3. Rapidly prepare amplicon libraries for illumina sequencers using ampliseq for illumina library preparation and index adapters. Kapa library quantification kit cycling parameters for the illumina eco reaction component final conc. This is dramatically faster than competing rna depletion kits, which require pretreatment protocols. Kapa biosystems kapa stranded rnaseq kit with riboerase. However, we do not know a priori the relative advantage of each method. Automated kapa hyperplus library preparation with the epmotion. All kits contain, at minimum, kapa sybr fast master mix 2x, kapa rt mix 50x, and dutp 10mm. Automated kapa hyperplus dna library preparation for. Kapa htp dna library preparation for illumina sequencing on the sciclone ngs workstation application. Kapa hifi pcr kits contain a novel dna polymerase, engineered for fast and versatile highfidelity pcr.
Fast and flexible processing of full or partial plates reproducible and accurate pipetting performance dispenses from 2 ul to 180 ul with 5 % cv 10 % accuracy installation qualification iq and operation qualification oq services are available versatile platform easily swappable head expands your throughput to 384 wells. The kit contains a novel dna polymeraseengineered via a process of molecular evolutionresulting in a unique enzyme specifically designed for realtime quantitative pcr qpcr using sybr green i dye chemistry. Kapa library quantification kapa library preparation kapa library amplification sybr fast the first enzyme engineered for qpcr. The standard curve is used to convert the average cq values for diluted library samples to concentration. Call, prioritize, and report on variants from one interface. Kapa library quantification kits coupled with the illumina. Targeted panel to investigate 40 dna genes, 29 rna fusion driver genes, and 5 gene expression levels associated with myeloid cancers. A performance evaluation of nextera xt and kapa hyperplus for rapid illumina library preparation of longrange mitogenome amplicons. The kit contains a novel dna polymerase engineered via a process of molecular. Ta cloning dna fragments generated with the kapa hifi pcr kit may be used directly for bluntend cloning, or cloning using restriction endonucleases. This standard operating procedure sop is for the quantification of next generation sequencing ngs libraries for illumina platforms on the applied biosystems abi viia7 instrument using quantstudio realtime pcr software v1. To find the right kit for your needs, use the selection tool below, or view a filterable list of library prep kits.
The kit uses a targeted enzymatic depletion method to remove ribosomal rna and thereby improve sequencing efficiency, the firm said. Illumina offers webinars and free online training courses to get the most out of your experiments and products. Kapa library quantification kit illumina platforms kr0405 v8. Kapa biosystems has launched its kapa stranded rnaseq kit with riboerase. This documents applies to kapa library amplification kits 07958978001 and 07958986001, kapa library amplification kits pcr mix only 07958943001. Kapa library quantification kits roche sequencing solutions. The result is a unique enzyme, specifically designed for qpcr using sybr green i dye chemistry.
Introduction illumina truseq nano dna library prep kit enables preparation of uniquely indexed dna libraries for single end and paired end sequencing figure 1. A performance evaluation of nextera xt and kapa hyperplus for. Seq library preparation kit illumina platforms, product codes kk8400. Kapa sybr fast onestep qrtpcr kit roche lightcycler 480 100 x 20 l reactions 100 x 20 l rxns,895 kk4701 kapa probe fast universal 2x qpcr master mix 1 ml 5,263 kk4702 kapa probe fast universal 2x qpcr master mix 5 ml 26,316. Optimal annealing temperatures are higher than in other. Kapa probe fast qpcr master mix 2x kits contain a readytouse cocktail containing all components except primers, probes, and template for fast cycling probebased qpcr. Ngs libraries prepared for sequencing on illumina and iontorrent platforms. Library quantification is performed by amplifying the set of six prediluted dna standards and appropriately diluted library samples by qpcr, using the kapa sybr fast qpcr master mix and primers based on the illumina p5 and p7 flow cell oligo sequences. It also provides an alternative to mrna enrichment and allows for the analysis of exonic regions, precursor mrnas, and long noncoding rnas. The workflow is fast and efficient, making it easy to assay tens of samples in parallel. Maestro kapa dna library preparation rev1 compatible with. Rad icycler qpcr kit 1 x 1ml illumina primer premix 10x 6 x 80ul illumina dna standards kapa biosystems kk4844.
Optimizing illumina nextgeneration sequencing library. Kapa library quantification kits comprise dna standards six 10fold dilutions and 10x primer premix, paired with kapa sybr fast qpcr kits to accurately quantify the number of amplifiable molecules in an illumina library. Sequencing library qpcr quantification guide 122363 c. Generating rna libraries from ultralowinput samples. Library quantification is performed by amplifying the set of six prediluted dna standards and diluted library samples by qpcr, using the kapa sybr fast qpcr master mix and primers targeting the illumina p5 and. Kits include kapa sybr fast qpcr master mix, a platformspecific library quantification primer premix.
Kapa hifi achieved a 100% success rate with higher yields up to 84% gc content. Kapa hifi pcr kit kit codes and components kr0368 v9. The kit contains a novel dna polymerase engineered via a process of molecular evolution. Kapa hifi dna polymerase has been designed for lowbias, highfidelity pcr, and is the reagent of choice for ngs library amplification2, 3, 4. Automate your roche and kapa library preparation are you looking for a solution to increase the throughput of your. The 452 bp kapa illumina dna standard consists of a linear dna fragment flanked by qpcr primer binding sites. Prior to cluster generation, amplified or nopcr libraries were quantified by qpcr using the sybr fast illumina library quantification kit kapa biosystems. Kapa library quantification kit universal from roche description kapa library quantification kits contain all the reagents needed for the accurate, reliable and reproducible qpcrbased quantification of ngs libraries prior to pooling for capture or flow cell amplification. Kapa biosystems kapa hyperplus and illumina s nextera xt kits are similarly priced and are both easily amenable to automation. The kapa htp library preparation kit kk8234 includes kapa hifi hotstart readymix 2x, a readytouse pcr mix comprising all the components for. Kapa sybr fast was engineered to amplify diverse dna fragments with similar. Kapa hyperprep kit collibri pcrfree ps dna library prep kit truseq dna pcrfree library prep kit kapa hyperprep kit 9 remove bias from genomic interpretation coriell na12878 samples were sequenced on an illumina novaseq 6000 system and reads normalized to 186m reads.
At illumina, our goal is to apply innovative technologies to the analysis of genetic variation and function, making studies possible that were not even imaginable just a few years ago. Kapa probe fast qpcr master mix 2x kit kk4701 100 x 20. Kapa library quantification kit kit codes and components. The average cq value for each dna standard is plotted against log 10 concentration in pm to generate a standard curve.
We then sequenced the libraries on the illumina platform. Kapa probe fast qpcr master mix 2x kit kk4701 100 x 20 l. Jan 07, 2019 we used the kapa stranded mrnaseq kit tradkapa to prepare libraries using the whole transcript method, and the lexogen quantseq 3mrnaseq library prep kitfwd 3lexo to prepare 3. Qiaseq fastselect rna removal kits qiagen online shop. Kits for ngs library preparation illumina offers an extensive assortment of easytouse nextgeneration sequencing library preparation kits for dna, rna, and epigenetic sequencing studies.
C ollibri ngs library preparation kits for highthroughput. Kapa sybr fast qpcr master mix 2x, available with various passive reference dyes refer to table 1 on page 6. Amplify fragments up to 15 kb from genomic dna or 18 kb from less complex targets. It is mission critical for us to deliver innovative, flexible, and scalable solutions to meet the needs of our customers. Library preparation kits optimized for illumina sequencers. Kk4202 kapa hrm fast pcr kit 5 ml 5 ml 36,842 kapa library preparation kits with standard pcr library next generation sequencing reagents kk8200 amplification illumina series 10 libraries 10 rxn 35,526 kk8201 kapa library preparation kits with standard pcr library. Please note that adaptors, primers, rrna depletion reagents and polya mrna isolation reagents are not included in the kit and are available separately. Product description kapa htp library preparation kit is designed for highthroughput library construction for illumina sequencing, starting from fragmented, doublestranded dna dsdna. Qiagen, eppendorf, illumina, and roche instruments do not require. Kapa hifi exhibits robust performance on targets with high gccontent. Kapa ltp library preparation kit illumina platforms product description the kapa ltp library preparation kit is a core kit for the construction of libraries for illumina sequencing, starting from sheared, doublestranded dna.
Kapa library amplification kit illumina platforms kk2620. Fast qpcr kits to accurately quantify the number of amplifiable molecules in an illumina library. Kapa library amplification kit illumina platforms kr0408 v7. Kapa htp library preparation kit illumina platforms. Improved library preparation with the nebnext ultra ii dna. A performance evaluation of nextera xt and kapa hyperplus. Kapa htp library preparation kit kit codes and components. Using kapa library preparation kits on the agilent ngs workstation yields highquality libraries for wholegenome sequencing on the illumina platform authors nguyet kong, kao thao, carol huang, and bart c. Kits include kapa sybr fast qpcr master mix, a platformspecific library quantification primer premix, and a prediluted. Weimer population health and reproduction department, school of veterinary medicine, university of california davis, davis, ca, usa maryke. Ngs library preparation illumina sequencing library prep. The surface provides excellent printing results, perfect. Kapa library quantification kit contains all reagents required for quantification of ngs libraries for illumina sequencing, including kapa sybr fast qpcr.
Kapa biosystems kapa hyperplus and illuminas nextera xt kits are similarly priced and are both easily amenable to automation. Kapa sybr fast dna polymerase has been engineered to perform. Kapa library quantification kits are optimized for the illumina genome analyzer, roche 454 titanium series. This new kit utilizes a fast, streamlined, automatable workflow for high yield production of superior quality libraries, with picogram to microgram input amounts of dna of varying quality. The kit provides all of the enzymes and reaction buffers required for the following steps. Quantitativerealtime pcr sybr fast the first enzyme. Flexibility to support manual and automated, highthroughput pipelines. Automated kapa library quantification kit with the epmotion 5075t.
Optimizing cluster density on illumina sequencing systems. To this end, we used two commercially available library preparation kits, the kapa stranded mrnaseq kit traditional. Library quantification primer plus kapa sybr fast illumina rox low 500 rkt. Kk8235 is available for pcrfree workflows and does not contain kapa. Kapa library quantification kit universal from roche. Kapa sybr fast qpcr master mix and primers based on the illumina p5 and p7 flow cell oligo. A panel of human amplicons with gc contents ranging from 66 84% was used to compare the performance of kapa hifi with that of an engineered polymerase with fusion technology and wildtype pfu. Our unique qiaseq fastselect procedure is compatible with qiagen, illumina, kapa and neb stranded library preparation kits and provides complete rrna removal in a single, 20minute inline step figure qiaseq fastselect rna removal workflow. Kapa sybr fast qpcr kit master mix 2x universal product description kapa sybr fast qpcr master mix is designed for high performance realtime pcr. The builtin analysis software streamlines data calculation. Kapa sybr fast was engineered to amplify diverse dna fragments with. Kapa sybr fast qpcr master mix 2x is designed for highperformance realtime pcr.
Using a qpcrbased solution, kapa library quantification kits provide accurate and reliable quantification of libraries prepared for sequencing on illumina and iontorrent platforms, across a wide range of library types, concentrations, fragment distributions and gc content. A fast, integrated workflow for a wide range of applications, from human wholegenome sequencing to amplicons, plasmids, and microbial species. Library quantification primer plus kapa sybr fast illumina lc480 500 rkt. Thus, a comprehensive comparison between the whole transcript and the 3.
Kapa htp library preparation kit for illumina platforms, v3. Kapa sybr fast onestep qrtpcr master mix 2x kit kr0393 v5. Kapa sybr fast qpcr master mix and primers based on the illumina p5 and p7 flow cell oligo sequences. Understanding library quantification assays for next. The 2x master mix contains kapa taq hotstart dna polymerase, dntps, mgcl 2, and stabilizers. Kits include kapa sybr fast qpcr master mix formulated with different. Free shipping through tcefs 5088561745 order must be submitted before 1pm for next day delivery email orders to. Sequencing library qpcr quantification guide illumina, inc.
Sequencing library qpcr quantification guide illumina. It is the optimal substrate for screen printing and also for any painting techniques. Kapa ltp library preparation kit kit codes and components. This document applies to kapa library quantification kits for illumina platforms 07960166001, 07960140001. Kapa sybr fast qpcr master mix 2x, available with various passive reference dyes. Automated kapa library quantification kit with the ep. Kapa hyper illumina nano nebnext ultra 1 ug 100 ng 10 ng robust performance with inputs from 1 ng 1 g fast, onetube workflow 2 3 hours with minimal handson time compatible with whole genome sequencing, targeted sequencing, pcrfree workflows, ffpe and low input samples contains kapa hifi and an optimized library.
Automated kapa hyperplus library preparation with the epmotion 5075t short protocol no. The qpcr was done following instructions provided by the kit manufacturer and run on the stepone realtime pcr system applied biosystems. Compare kapa hyper prep kitillumina, with kapa library amplification primer mix 10x from roche on. A comparison between whole transcript and 3 rna sequencing. Kapa library quantification kit contains all reagents required for quantification of ngs libraries for illumina sequencing, including kapa sybr fast qpcr master mix 2x, a primer premix 10x containing two primers based on illumina p5 and p7 oligo sequences primer 1. Each nextera dna flex library prep kit requires one of the following nextera dna flex index kits to complete the protocol, regardless of the sample. Kapa hyper prep kitillumina, with kapa library amplification. Automated kapa library quantification kit with the. Flexibility to support manual and automated highthroughput pipelines as well as. Illumina libraries with an average fragment length of up to. However when moving toward the adoption of ngs for forensic casework, a balance must be struck between throughput and coverage.
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