qPCR standard curve FAIL


Several months ago I agreed to give a talk next Tuesday. At the time it seemed a given that by then I would have some beautiful qPCR data. And then my minus 80 freezer died, taking with it all of my vital tissue samples, which took a further 6 weeks to regrow before I could even think about RNA extractions and DNase treating and cDNA synthesis and qPCR.

This has left me with a matter of days to finish round one of qPCR in time for my talk. (And given that I’m moving house over the weekend, it’s not the greatest time for late night lab sessions.) Today I ran a standard curve to check how little DNA I could get away with using and still get a decent signal.

The three beautiful linear lines are my house keeping genes (which are used to normalise data). The stupid flat thing filling me with rage is meant to be my gene. Or would be, if my primers worked or weren’t contaminated or something. I don’t even know.

Massive lab fail and almost certainly too late to do anything about it.

Someone hand me an undergrad because right now nothing less than blood sacrifice will satisfy the vengeful lab gods…

Advertisements

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s