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Old 12-18-2015, 03:03 PM   #1
DRYTCYV
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Default MiSeq Failed Run

Hello,

I got this:

"Your run xxxxxxxx on instrument xxxxxxx recently encountered the following error during sequencing:

<Internal Illumina Command Exception>"

Any ideas?

Thanks
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Old 12-18-2015, 04:59 PM   #2
GenoMax
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Don't mean to state the obvious but have you contacted tech support after taking a screen shot?
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Old 12-22-2015, 02:53 AM   #3
DRYTCYV
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Quote:
Originally Posted by GenoMax View Post
Don't mean to state the obvious but have you contacted tech support after taking a screen shot?
MiSeq Run started: 20:30 (Friday)
Error msg : 21:30

Who to call?

A) Illumina techsupport
B) SEQanswers amazing tech support (and sometimes the only tech support)
C) Ghostbusters

The correct option : B

The error msg was just from the e-mail, when something bad happened, but in the machine I got this one:

"No usable signal found in the images; it is possible that clustering has failed"

Possible Cause: NaOH concentration too high
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Old 12-22-2015, 06:26 AM   #4
Jessica_L
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If you spiked in PhiX and you're relatively certain your libraries should have been fine, I think NaOH is at least worth a look. Do you normally check the pH of your 0.2N NaOH before using it to denature? I used to not do it and had so many runs fail because of bad NaOH from Illumina (came in our HiSeq kits) that I started checking it every single time.

Illumina tech support will ask you to run some of the machine tests (volumetric tests, etc.) so you can do some of that preemptively. I'd still contact Illumina because you should be able to get a replacement kit if everything else checks out.

I've seen cluster generation failures when there's just not enough signal (i.e. the loading concentration of the library is too low) so it might be worth double checking any quantitation measurements that were made, too.
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Old 12-22-2015, 06:27 AM   #5
GenoMax
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So was that the cause of the failure? It sounds like you were able to re-run with correct concentration of NaOH?
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