Interpretation of BioAnalyzer results?

[aspencoyle]

Alright, I extracted DNA from 3 Tanner crab samples from 2005 and then ran them on the Qubit and BioAnalyzer. Goal was to assess DNA quality from these archived samples and determine if a large-scale project is feasible.

I'm not entirely sure how to interpret these BioAnalyzer results - any feedback on what this looks like in terms of quality?

Links to:

Qubit results

Notes on extraction

And BioAnalyzer results are below. Note: The 4th sample ran on the BioAnalyzer was a blank meant to test whether the gel (expired) was still good.

[kubu4]

Looks like the markers weren't run or didn't work. Will need to be re-run.

I wouldn't use expired reagents for Bioanalyzer.

Expired chips are okay, though.

[aspencoyle]

Gotcha, how can you tell the marker didn't work? Trying to learn all this! And do you know if we have fresh reagents (for DNA specifically)?

[kubu4]

I can tell the markers didn't work because they're not present in any of the lanes. The marker serves two purposes:

• positive control
• time reference

There should be a small marker and a large marker in all lanes. These are represented as tall narrow peaks very early (small marker) in the measurement time and very late (large marker) in the measurement time.

I'd doubt we have any usable Bionalyzer reagents. Expiration date is usually about 6 months after receipt, and is indicated on the box.

[kubu4]

I'll order a new set of reagents Monday.

[aspencoyle]

Thanks a bunch, Sam! Can you let me know when they get in?

[kubu4]

@afcoyle Kit is scheduled for delivery today. FedEx tracking number: 271310528357