The NEBNext® Single Cell/Low Input cDNA Synthesis & Amplification Module uses a template switching method to generate full length cDNAs directly from single cells or 2 pg – 200 ng RNA. This unique workflow enables generation of high quality sequencing data from a broad range of inputs, and superior transcript detection, while providing reliably consistent performance.
Features:
The highest yields from single cells, or 2 pg - 200 ng RNA
Input can be cultured or primary cells, or total RNA
Single-tube protocol from cell lysis to cDNA
Fast, streamlined, automation-friendly workflow, with minimal handling steps and hands-on time
Download extensive performance data in our technical note.
Figure 1: NEBNext Single Cell/Low Input workflow
Figure 2: Higher cDNA yields with the NEBNext Single Cell/ Low Input cDNA Synthesis & Amplification Module
Sequencing libraries were generated from HeLa, Jurkat and M1 single cells, or 10 pg of Universal Human Reference (UHR) RNA (Agilent® #740000) with recommended amounts of ERCC RNA Spike-In Mix I (Thermo Fisher Scientific® #4456740). The NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module or the SMART-Seq® v4 Ultra® Low Input RNA Kit for Sequencing (Clontech® # 634891) were used. cDNA libraries were amplified using the number of PCR cycles shown. Error bars indicate standard deviation for 8-21 replicates.
核糖核酸酶A是内切核糖核酸酶,可特异地攻击RNA上嘧啶残基的3'端,切割与相邻核苷酸形成的磷酸二酯键。反应终产物是嘧啶3'磷酸及末端带嘧啶3'磷酸的寡核苷酸。无辅因子及二价阳离子存在时,核糖核酸酶A的作用可被胎盘RNA酶抑制剂(B1ackburn et al.1977)或氧钒—核糖核苷复合物(Puskas et al.1982)所抑制。核糖核酸酶A用途生化研究,测定核酸的结构RNase 保护检测去除非... 查看更多>
DNase I 是一种核酸内切酶,降解双链或单链DNA,产生5-磷酸末端的单核苷酸及寡核苷酸,在Mg2+存在时,DNase I独立地作用每条DNA链,切割位点是随机分布,在Mn2+存在下,DNase I 作用于DNA双链的大致同一位置,产生钝末端或具1-2个核苷酸突起的DNA片段。来源:牛胰贮存条件:4℃应用:1、用切口平移法进行放射性标记时,可用DNase I在双链DNA上产生随机切口;2、在进行亚硫酸氢盐介导的诱变前,可用DNase... 查看更多>