Hi,
I have a really burning project idea I want to pursue. The problem is my PI wont let me. Hence I am thinking of financing the project out of my own pocket.
In order for me not to ``waste`` any time doing my own stuff in his lab, I might have to ``secretly`` outsource nearly every experimental procedure. I was looking towards websites like scienceExchange for providers.
What I am planning to do is a simple CRISPR HDR GFP knock in in some cancer cell lines. I just need to clone the sgRNA CRISPR vector, clone the HDR GFP vector, transfect and measure the integration efficiency.
So far, I have saved up $20,000 of my own money.
Do you think that is sufficient to outsource everything? If it is, then how many replicates on different cell lines can I afford?
If that isnt, then how much more do I need?
Thanks for your help.
You will need a lot more than $20k. Generating the KO lines is the "easy" part. The time-consuming and costly part will be the phenotypic characterization. This is more like a $100k to $200k project. It's a bit like genome sequencing: $1k for a genome; $50-100k for the bioinformatics...
Hi Mark,
I dont need any phenotypic characterisation. Im developing a new method to increase HDR efficiency in cell lines.
I just want to measure how well cells are labbeled with GFP (i.e how many % GFP positive) using the method that Im developing. SImple fluorescent microscopy will do.
In other words, how far does $20k go when it comes just to knocking in cell lines?
It's unlikely that detection of GFP fluorescence will provide info on HDR because random insertion of promoter-less GFP vectors frequently results in expression due to integration downstream of sequences with promoter activity.
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