bacterial RNAseq using TruSeq Lib prep

thiNGS · 03-09-2018, 05:08 AM

Hi all. I am in the process of starting RNAseq from bacterial samples and I am trying to identify the best strategy to do it. Given that bacterial RNA is not poly-adenylated, a random hexamer-based system is essential. Has anyone used the TruSeq stranded library prep kit for bacterial RNA? Or do you have a good kit to recommend? The samples will be run on the MiSeq or NextSeq systems. Any advice to guide me through the magical world of RNAseq will be much appreciated

huguesparri · 03-13-2018, 02:15 AM

We used the Truseq kit succesfully on Bacteria. 2 things are essential:
- DNAse treatment of your RNA.
- Good depletion efficiency for the ribosomal RNA. Ribozero kit are usually doing well.

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03-09-2018, 05:08 AM	#1
thiNGS Member Location: Netherlands Join Date: Sep 2014 Posts: 24	bacterial RNAseq using TruSeq Lib prep Hi all. I am in the process of starting RNAseq from bacterial samples and I am trying to identify the best strategy to do it. Given that bacterial RNA is not poly-adenylated, a random hexamer-based system is essential. Has anyone used the TruSeq stranded library prep kit for bacterial RNA? Or do you have a good kit to recommend? The samples will be run on the MiSeq or NextSeq systems. Any advice to guide me through the magical world of RNAseq will be much appreciated

03-13-2018, 02:15 AM	#2
huguesparri Member Location: Montpellier (France) Join Date: May 2008 Posts: 93	We used the Truseq kit succesfully on Bacteria. 2 things are essential: - DNAse treatment of your RNA. - Good depletion efficiency for the ribosomal RNA. Ribozero kit are usually doing well.