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RNaseZapTM RNase Decontamination Solution

Company: Ambion
Catalog#: AM9780
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Using RNA Sequencing and Spike-in RNAs to Measure Intracellular Abundance of lncRNAs and mRNAs
Author:
Date:
2020-10-05
[Abstract]  Long noncoding RNAs (lncRNAs) play essential roles in normal physiology and in disease but their mechanisms of action can be challenging to identify. For mechanistic studies, it is often useful to know a lncRNA’s intracellular abundance, i.e., approximately how many molecules of the lncRNA are present in a typical cell of a cell-type of interest. At least two approaches have been used to approximate lncRNA intracellular abundance: single-molecule sensitivity RNA fluorescence in situ hybridization (smFISH) and single-gene, calibrated reverse-transcription followed by quantitative PCR (RT-qPCR). However, like all experimental approaches, these methods have their limitations. smFISH, when analyzed using diffraction-limited microscopy, can underestimate intracellular ... [摘要]  [摘要]长非编码RNA(lncRNA)在正常生理和疾病中起着至关重要的作用,但其作用机理可能难以鉴定。对于机理研究,了解lncRNA的胞内丰度(即在目标细胞类型的典型细胞中大约存在多少个lncRNA分子)通常很有用。至少两种方法已用于估算lncRNA细胞内丰度:单分子敏感性RNA荧光原位杂交(smFISH ...

Whole-genome Identification of Transcriptional Start Sites by Differential RNA-seq in Bacteria
Author:
Date:
2020-09-20
[Abstract]  Gene transcription in bacteria often starts some nucleotides upstream of the start codon. Identifying the specific Transcriptional Start Site (TSS) is essential for genetic manipulation, as in many cases upstream of the start codon there are sequence elements that are involved in gene expression regulation. Taken into account the classical gene structure, we are able to identify two kinds of transcriptional start site: primary and secondary. A primary transcriptional start site is located some nucleotides upstream of the translational start site, while a secondary transcriptional start site is located within the gene encoding sequence.

Here, we present a step by step protocol for genome-wide transcriptional start sites determination by differential RNA-sequencing (dRNA-seq) ...
[摘要]  [摘要] 细菌中的基因转录通常起始于起始密码子上游的一些核苷酸。识别SPE cific Ť ranscriptional 小号挞小号ITE (TSS)为遗传操作必需的,因为在许多情况下,起始密码子上游有中涉及的基因表达调控序列元件。考虑到经典的基因结构,我们能够鉴定出两种转录起始位点:一级和二级。主要转录起始位点位于翻译起始位点上游的一些核苷酸上,而次要转录起始位点位于基因编码序列内。

这里,我们提出一步步协议全基因组吨ranscriptional 小号馅饼小号ITES d etermination通过差RNA测序(DRNA 使用肠道病原体-SEQ)福氏痢疾杆菌血清型菌株5A作为M90T模型。但是,该方法可以用于选择的任何其他细菌物种。第一步,使用热酚法从细菌培养物中纯化总RNA。核糖体RNA(rRNA)是使用商业试剂盒通过杂交探针特异性去除的。然后准备一个富含5'- 一磷酸依赖性核酸外切酶(TEX)处理的,富含初级转录本的RNA文库,用于与未进行TEX处理的文库进行比较,然后连接已知序列的RNA接头衔接子,从而确定具有单核苷酸精度的TSS。最后,对RNA进行处理以制备Illumina测序文库,并按购买的服务进行测序。通过内部生物信息学分析鉴定TSS。

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