Hello,
I want to look into MiSeq technique for gene expression analysis. I would like to know if there are any tips, guidelines or suggestions. Also if there are other ways to choose a housekeeping gene for normalization.
Anything to get started would be greatly appreciated. Thanks
Please look into
a) https://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-0881-8
b) http://www.cell.com/trends/plant-science/abstract/S1360-1385(13)00024-1
Hi Sana,
I have been using the MiSeq for Targeted RNASeq assays. With these types of assays you can sequence/quantify hundreds of genes at the same time per sample, with multiplexing. I have been using Qiagens targeted RNASeq panels, but you can also customize specific genes panels. It is highly quantitative due to the use of unique molecular bar-codes per transcript/per gene. The normalization is based on 10 housekeeping genes, but you can also include your own housekeeping genes if you choose so. QC such as checks for genomic DNA contamination is included with the assay, and the bioinformatics is already taken care off through Qiagen's algorithms.
Hope that helps.
Mahmood
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