货号：4475936

品牌：ThermoFisher /Invitrogen/ Gibco/ ABI

规格：EACH

目录价：￥20828.00

市场价格：￥18745.20

会员价格：￥18745.20

金山科研平台，产品价格货期咨询微信：jinshanbio 描述 The Ion Total RNA-Seq Kit v2 includes the reagents needed to prepare representative cDNA libraries for strand-specific RNA sequencing on the Ion Proton™ or the Ion Personal Genome Machine® (PGM™) systems. The Ion Proton™ System is ideally suited for sequencing the whole transcriptome (ribosomal RNA depleted or polyA), while the Ion PGM™ System is suitable for small RNA, as well as viral and bacterial transcriptomes. Either small RNA (such as microRNA) or whole transcriptome RNA samples can be prepared for next-generation sequencing using the Ion Total RNA-Seq Kit v2. Version 2 of the Ion Total RNA-Seq Kit is an improvement over the first generation kit. The gel size selection step in the small RNA workflow has been replaced with a magnetic bead-based method. Magnetic bead-based purification replaces all of the filter cleanup steps and the total reaction time has been reduced to 6 hours.Additional Features of the New Ion Total RNA-Seq Kit v2:• Greater accuracy—SuperScript® VILO™ and Platinum® PCR SuperMix High Fidelity added for highest template fidelity• Barcode compatible—works with Ion Xpress™ RNA-Seq Barcode 01-16 Kit for multiplexing• Automation friendly—magnetic bead-based purification simplifies automation of library constructionAs with the previous kit, the Ion Total RNA-Seq Kit v2:• Preserves strand information—all mapped reads are aligned in the direction of transcription relative to the chromosomal strand• Allows you to choose your workflow—interrogate either small RNA or the whole transcriptome• Allows you to analyze any type of RNA—supports small RNA, rRNA depleted total RNA, and poly(A) RNAThe Ion Total RNA-Seq Kit v2 is designed to make cDNA library preparation for the Ion Proton™ and PGM™ systems fast and flexible. It can be used to generate a representative cDNA library, flanked by specific sequences necessary for sequencing, from any type of RNA sample.Designed as a complete solution with a common workflow, the Ion Total RNA-Seq Kit v2 combines optimized reagents and protocols for discovery of small RNAs and isoforms, coding RNA, noncoding RNA, and alternative splice variants.Small RNA AnalysisDuring small RNA library construction, the 3' and 5' adaptors are attached directionally and simultaneously. As a by-product of this step, an adaptor:adaptor product may be formed without an RNA insert. This byproduct will amplify during first strand synthesis and PCR. If not removed, > 50% of the reads will be the adaptor dimer. Historically, the only way to separate the adaptor dimer from the wanted small RNA containing library fragments has been through gel size selection. The Ion Total RNA-Seq Kit v2 uses proprietary technology to inhibit cDNA synthesis of the adaptor byproduct, thus allowing cDNA separation with magnetic bead-based technologies. Total reaction time has been reduced from 1.5 days to approximately 6 hours. A separate Magnetic Bead Cleanup Module is included with the kit. Additional modules may be purchased separately.Start with total RNA containing as little as 5–100 ng of miRNA, or RNA enriched for small RNA containing 1–100 ng of miRNA. The small RNA protocol provides guidance on whether to start with total RNA or RNA enriched for small RNA, based on the small RNA content of your sample. Small RNA enrichment protocols are also provided.Whole Transcriptome AnalysisThe whole transcriptome protocol enables construction of strand-specific libraries in approximately 5 hours. Starting with as little as 100 ng of total RNA, construct a library from 1 ng of poly(A) RNA or 25 ng of rRNA-depleted RNA following the RNA enrichment and library generation protocols provided in the manual. Because the libraries are not limited to cDNA derived only from poly(A) RNA, Ion Total RNA-Seq Kit libraries support a more thorough investigation of transcriptome complexity, capable of characterizing known and undocumented transcripts, including alternative splice variants, fusion transcripts, and SNPs.Preserve Strand InformationUnlike methods that ligate adapters to double-stranded cDNA, the Ion Total RNA-Seq Kit v2 utilizes proprietary Ambion® technology to attach the adapters in a directional manner that preserves strand information in the resulting libraries. In addition, both the 3' and 5' adapters are attached simultaneously, reducing ligation and clean-up steps.Preserving strand orientation during library construction helps enable more accurate determination of the structure and expression level of transcripts, and can aid in the discovery of novel transcription regions from both the positive and negative genomic strands.The Ion Total RNA-Seq Kit v2 is designed to create RNA libraries from up to 12 samples for small RNA or whole transcriptome sequencing on the Ion Proton™ or PGM™ systems. 规格

Platform:	Ion PGM™ System
Workflow Step:	Library Generation
Product Size:	12 reactions

内容及储存 Ion RNA-Seq Core Kit v2Store at -20°C to room temperature:2 x 1.75 ml Nuclease-free WaterStore at -20°C:1 x 20 µL 10X RNase III Reaction Buffer1 x 20 µL RNase III1 x 40 µL Hybridization Solution1 x 150 µL 2X Ligation Buffer1 x 30 µL Ligation Enzyme Mix1 x 30 µL 2.5 mM dNTP Mix1 x 56 µL 10X SuperScript® III Enzyme Mix1 x 900 µL Platinum® PCR SuperMix High Fidelity1 x 50 µL WT Control RNA (1 µg/µL HeLa total RNA)1 x 10 µL Small RNA Control (1 µg/µL human placenta total RNA)1 x 10 µL Small RNA Control (1 µg/µL human placenta total RNA)Ion RNA-Seq Primer Set v2 (Cat. No. 4474810)Store at -20°C:1 x 30 µL Ion Adaptor Mix v21 x 104 µL Ion RT Primer v21 x 20 µL Ion 5' PCR Primer v21 x 20 µL Ion 3' PCR Primer v2