analysis of small bacterial RNA on MiSeq
Hello everyone,
First time posting, please be gentle.
We need to analyze small regulatory RNAs in Pseudomonas. The RNAs are 50 to 450 bp long, and have the 5'P-P-P and 3'OH structure. My idea was to use a Qiagen miRNeasy kit, with the small RNA enrichment protocol (in the appendix). The resulting fraction should be enriched for the correct-sized RNA. The question then is how to prepare a library for MiSeq. I looked into the Illumina TruSeq small RNA method, and did not like it at all. My preference would be to use something like the SMARTer Ultra Low RNA Kit for Illumina Sequencing from Clontech, without size selection.
Does anyone have any experience with that kit, or can offer alternative ideas or suggestions? Much appreciated.
First time posting, please be gentle.
We need to analyze small regulatory RNAs in Pseudomonas. The RNAs are 50 to 450 bp long, and have the 5'P-P-P and 3'OH structure. My idea was to use a Qiagen miRNeasy kit, with the small RNA enrichment protocol (in the appendix). The resulting fraction should be enriched for the correct-sized RNA. The question then is how to prepare a library for MiSeq. I looked into the Illumina TruSeq small RNA method, and did not like it at all. My preference would be to use something like the SMARTer Ultra Low RNA Kit for Illumina Sequencing from Clontech, without size selection.
Does anyone have any experience with that kit, or can offer alternative ideas or suggestions? Much appreciated.
