I'd like to know that what are the different ways to know/identify whether a particular Gene is expressed or not ?
Few points from my side are :
1) identifying it's corresponding m-RNA transcripts level.
2) identifying the protein that was produced by the expression of that particular Gene.
Any other points ?
or go to the GTEX portal where you'll get informations on expression levels, type of cells, transcripts...and much more.
https://gtexportal.org/home/
fred
Hi Vikram Kumar
First you have to keep in mind that mRNA might not reflect the corresponding protein level and vice versa. So it is better to elaborate what do you mean by the gene expression and your suggestion to detect it by western blot...etc? for example, some protein may aggregate without an overexpression of the corresponding gene like p-a-synuclein.
So, if you would like to make a conclusion only over the expression then yes you can use PCR, but you have to take in your consideration the type of PCR that would give you a hand.
On the other hand, if your target is to detect the protein level, then WB, ELISA, Flowcytometry ..etc would be the solution.
Dear Vikram Kumar
You can also tag the protein of interest with a reporter gene like fluorescence protein (GFP/mCherry) or with an epitope tag like HA-tag, FLAG-tag, 6xHis-tag etc. If it is a fluorescence protein tag, you can primarily visualize under fluorescence microscope followed by detection with western blot. Epitope tagging is easy to identify as the respective antibody has high affinity with the tag. However, the selection of an appropriate linker between the protein of interest and the tag is a crucial step.
Best wishes.