Ion Total RNA-Seq 试剂盒 v2
Ion Total RNA-Seq 试剂盒 v2
Ion Torrent™

Ion Total RNA-Seq 试剂盒 v2

Ion Total RNA-Seq 试剂盒 v2 包括在 Ion GeneStudio S5、Ion了解更多信息
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货号反应次数
447978912 或 48
447593612 或 48
货号 4479789
价格(CNY)
-
反应次数:
12 或 48
Ion Total RNA-Seq 试剂盒 v2 包括在 Ion GeneStudio S5、Ion Proton 或 Ion Personal Genome Machine (PGM) 系统上为链特异性 RNA 测序制备代表性 cDNA 文库所需的试剂。Ion GeneStudio S5 和 Ion Proton 系统是适用于全转录组(去除核糖体 RNA 或 polyA)测序的理想选择,而 Ion GeneStudio S5 和 Ion PGM 系统适用于病毒和细菌转录组。Ion Total RNA-Seq 试剂盒版本2是第一代试剂盒的改进版本。磁珠法纯化替代了所有过滤纯化步骤,总反应时间减少到6小时。

新 Ion Total RNA-Seq 试剂盒 v2 的其他特点:

•准确度更高 — 添加 SuperScript VILO 和 Platinum PCR SuperMix High Fidelity 以达到极高的模板保真度
•兼容条形码 — 与用于多通路检测的 Ion Xpress RNA-Seq 条形码01-16试剂盒配合使用
•适用于自动化 — 磁珠法纯化可简化文库构建的自动化

与之前试剂盒一样,Ion Total RNA-Seq 试剂盒 v2:

•保留链的信息 — 所有图谱读段与染色体链相关的转录方向一致
•使您可以分析不同类型的 RNA — 支持去除 rRNA 的总 RNA 和 poly(A) RNA

Ion Total RNA-Seq 试剂盒 v2 设计用于使 cDNA 文库制备快速灵活。它可用于通过任何类型的 RNA 样品生成代表性 cDNA 文库,以测序所需要的特定序列为侧翼。

通过设计为包含普通工作流程的完整解决方案,Ion Total RNA-Seq 试剂盒 v2 结合了优化的试剂和方案,可发现编码 RNA、非编码 RNA 和选择性剪接变体。

全转录组分析
全转录组方案可确保在约5小时内构建链特异性文库。按照手册中提供的 RNA 富集和文库生成方案,从低至 100 ng 的总 RNA 开始,使用 1 ng poly(A) RNA 或 25 ng 去除 rRNA 的总 RNA 构建一个文库。由于文库不限于仅来源于 poly(A) RNA 的 cDNA,Ion Total RNA-Seq 试剂盒文库支持对转录组复杂性进行更全面的研究,能够表征已知和未记录的转录本,包括选择性剪接变体、融合转录本和 SNP。

保留链信息
与连接接头至双链 cDNA 的方法不同,Ion Total RNA-Seq 试剂盒v2使用专有的 Ambion 技术以定向方式连接接头,以保留生成的文库中的链信息。此外,同时连接 3' 和 5' 接头,减少连接和纯化步骤。

在文库构建期间保持链定向有助于更精确测定转录本的结构和表达水平,并有助于从阳性和阴性基因组链中发现新型转录区域。

Ion Total RNA-Seq 试剂盒 v2 设计用于利用至多48份样品创建 RNA 文库,以在 Ion GeneStudio S5、Ion Proton 或 Ion PGM 系统上进行全转录组测序。
仅供科研使用。不可用于诊断程序。
规格
适用于(设备)Ion GeneStudio™ S5、Ion PGM™、Ion Proton™ 系统
cDNA 文库
反应次数12 或 48
产品线Ion Torrent™
数量48 reactions
测序类型转录组测序
原始材料量对于 RNA(去除 rRNA):1-100 ng mRNA
对于全转录组:≥100 ng 总 RNA
工作流程步骤NGS 文库制备
样品类型RNA
Unit SizeEach

常见问题解答 (FAQ)

What is an indicator of a successful RNA library?

Generally, a successful Ion RNA-Seq library will be within the recommended size range for sequencing on the Ion PGM or Proton systems and the sequencing data will have a high (or expected) percentage of on-target reads.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

I am new to RNA-Seq. Where can I find information on how to approach the sequencing data analysis?

Please visit the Transcriptome Sequencing by Ion Torrent Sequencing page (https://www.thermofisher.com/us/en/home/life-science/sequencing/rna-sequencing/transcriptome-sequencing/transcriptome-sequencing-ion-torrent-next-generation-sequencing.html) for information on RNA-Seq data analysis using Torrent Suite Software, or 3rd party software providers.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Why do I see the sequence GGCCAAGGCG at the beginning of the majority of the reads in my sequencing data analysis?

The GGCCAAGGCG sequence is part of the RNA library adapter sequence. This sequence can be automatically trimmed by the software by selecting the “RNA_Barcode_None” option under the Barcodes menu. The run can be reanalyzed with the correct barcode settings to trim the adapter sequence. In the future, the “RNA_Barcode_None” option should be selected in the run plan or selected on the instrument prior to starting the sequencing run.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Which Ion platform and chip should I use for whole transcriptome sequencing?

RNA-Seq relies on a random sampling strategy in which most sequencing reads will identify abundant transcripts with an increasing number of reads needed to identify lower abundant transcripts and/or low levels of gene expression. While depletion and purification strategies are available from Thermo Fisher Scientific to remove unwanted abundant transcripts and increase sequencing power, transcript detection sensitivity is a function of mapped reads. Accordingly, the complexity of the genome under investigation should be taken into account when planning an RNA-Seq experiment. Applications that require a small number of reads, such as low multiplexed gene expression, can be performed on the Ion 318 Chip Using the PGM platform whereas, human/mammalian transcriptomes and highy-multiplexed gene expression profiling should be performed on the GeneStudio platforms, using the higher throughput chips. For more information, please see the Transcriptome Sequencing by Ion Torrent Sequencing page (https://www.thermofisher.com/us/en/home/life-science/sequencing/rna-sequencing/transcriptome-sequencing/transcriptome-sequencing-ion-torrent-next-generation-sequencing.html).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

The Ion Total RNA-Seq Kit v2 utilizes RNase III for RNA fragmentation in the whole transcriptome library preparation workflow. Are there other forms of fragmentation?

Another method is chemical fragmentation (not part of the protocol), which is useful for quantifying exon levels, novel splicing identification, and fusion gene discovery, and provides more uniform coverage for viral and bacterial transcripts. Chemical fragmentation is more randomized than RNase III fragmentation, but can be difficult to optimize. Chemically fragmented RNA fragments require repair with T4 polynucleotide kinase (not included in kit) before proceeding into ligation.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.