
Strand-specific Illumina sequencing libraries from low RNA inputs of any quality
The SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian provides a complete solution for sensitive and streamlined total RNA analysis. The kit is designed to work with a wide range of input amounts (10–100 ng) of total RNA of any quality. A variety of human, rat, and mouse samples can be used, including total RNA from FFPE material, tissue samples, etc. The kit provides whole transcriptome analysis with accurate measurement of strand orientation, unbiased coverage, high transcript mapping, and gene counts.
Strand-specific Illumina sequencing libraries from low RNA inputs of any quality
The SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian provides a complete solution for sensitive and streamlined total RNA analysis. The kit is designed to work with a wide range of input amounts (10–100 ng) of total RNA of any quality. A variety of human, rat, and mouse samples can be used, including total RNA from FFPE material, tissue samples, etc. The kit provides whole transcriptome analysis with accurate measurement of strand orientation, unbiased coverage, high transcript mapping, and gene counts.
This low-input kit contains the Illumina Indexing Primer Set, and the entire protocol—from rRNA depletion through the creation of Illumina-specific, indexed RNA-seq libraries—takes only about 5 hours.
Alternate kits for other input amounts:
- For 250 pg–10 ng of total RNA, we recommend the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian
- For 100 ng–1 µg of total RNA, we recommend the SMARTer Stranded Total RNA Sample Prep Kit - HI Mammalian
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对于新手来说购买反转录试剂盒是比较理想的,买一个kit看看说明书操作就可以了。推荐两款kit TAKARA和HaiGene,这两款试剂盒都能对RNA样品中的gDNA有效去除,因此对RNA质量的要求不高。TAKARA的RR047A操作方便,反转录温度为37℃,对于大多数试验来讲是满足要求的。HaiGene的D0401操作要多一个步骤,但其反转录温度是55℃(耐高温的反转录酶),提高了反转录温度使得高GC含量、复杂模板、长mRNA的模板都能有效反转录,因此其反转录效率更高,更能够获得样本中基因的真实表达量。如果后续试验是RealTime PCR、ORF克隆、高GC含量、或者你的待研究基因结构复杂程度未知,还是选用耐高温的反转录酶更理想。对于反转录高手来说,直接购买反转录酶、再购买Rnase Inhibitor自己配制反转录体系就可以了。对于样本量大的课题组来讲,相对还是比较经济的。选择反转录酶时仅需要考虑是否需要耐高温的酶,来克服目的基因的复杂结构就可以了。PROMEGA、TAKARA、HaiGene、 TRANSGEN等品牌的反转录酶性价比还都是不错的。Life、NEB的也不错,不过性价比一般,不一一解释了。

