The RNA-Seq protocol described here yields strand-specific transcriptome libraries without poly(A) selection, which provide approximately 90% mappable sequences. Typically, more than 85% of mapped reads correspond to protein-coding genes and only 6% derive from non-coding RNAs. The protocol has ...
This unit presents streamlined workflows for generating strand-specific RNA-seq libraries from 10ng to 1g total RNA, representing a minimum of 1000 cells, in less than 7hr with minimal carryover rRNA. These methods allow scientists to evaluate the expression of all transcripts, including non-poly...
Strand-specific RNA sequencing (ssRNA-Seq) can overcome these limitations and as such is better suited for genome annotation, de novo transcriptome assembly and accurate digital gene expression analysis. This protocol describes a simple and robust method to generate ssRNA-Seq libraries for the ...
To determine whether R-loops form at PREs, we carried out two biological replicates of strand-specific DNA-RNA Immunoprecipitation followed by next generation sequencing (DRIP-seq) in Drosophila embryos (2–6 and 10–14 hour (H)) and in S2 cells (Fig. 1, Supplementary Fig. 1). DRIP-se...
They demonstrated that RIs contain a ssDNA-specific nuclease-resistant Y arc, suggesting that the long single-stranded stretches of the parental heavy strand predicted by the strand-displacement model were absent. Instead, the authors proposed that the RIs observed in their analysis resulted from a ...
Tn5 tagmentation of the RNA/cDNA hybrid [23]. Regardless of the specific protocol, the inevitable RT/SSS with a limited efficiency in existing scRNA-seq methods ultimately compromises the single-molecule capture efficiency of the original RNA molecules. Low RNA capture efficiency can lead to ...
Over-expression of ATH leads to aberrant R-loop accumulation, which can be attenuated by the over-expression of AtRNH1C synchronously. Strand-specific DNA damage sequencing revealed that transcription-replication competition could introduce single-strand DNA breaks near the end of the transcription ...
First strand cDNA synthesized by random hexamer priming on RNA showed consistent position and nucleotide-specific mismatch errors in the first seven nucleotides. The mismatch errors found in both datasets are consistent in distribution and thermodynamically stable mismatches are more common. This strongly ...
SureSelect Automated Strand-Specific RNA Library Prep Automated Poly-A Selection and Strand-Specific mRNA Library Preparation for the Illumina Platform Protocol Version F0, June 2020 SureSelect platform manufactured with Agilent SurePrint Technology For Research Use Only. Not for use in diagnostic ...
gDNA remover 可以有效的去除 RNA 提取过程中残留的基因组DNA,处理后的样品可以直接用于后续实验。优化的反应体系(5×Buffer (with primer))中预混了Random Primer 和 Oligo(dT)18 Primer,简化了加样操作步骤,避免了操作误差和污染的风险;本产品可以从极低量的总 RNA 或 poly(A) mRNA 合成第一链 cDNA,合成的...