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Transcriptional noise chadn737 RNA Sequencing 9 03-01-2011 10:28 AM

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Old 06-10-2016, 12:17 PM   #1
soleulloa
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Default Z Motor: ADC noise which exceeds ADCNoiseErrorLimit

Hi all,
Yesterday a run in MiSeq was stopped showing the next message:

Z Motor: ADC noise is 3592 which exceeds ADCNoiseErrorLimit 3277. numSamples=10x8, avg=31258, posNoise=1906, negNoise=3592

Density cluster was detected and it was 1266 K/mm2.


Anyone have seen that error??
What does ir means?

Regards!!
Soledad
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Old 06-10-2016, 12:34 PM   #2
kmcarr
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Quote:
Originally Posted by soleulloa View Post
Hi all,
Yesterday a run in MiSeq was stopped showing the next message:

Z Motor: ADC noise is 3592 which exceeds ADCNoiseErrorLimit 3277. numSamples=10x8, avg=31258, posNoise=1906, negNoise=3592

Density cluster was detected and it was 1266 K/mm2.


Anyone have seen that error??
What does ir means?

Regards!!
Soledad
This is a hardware error on your MiSeq. You need to contact Illumina Technical Support.
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Old 06-10-2016, 09:58 PM   #3
soleulloa
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Hi kmcarr,
Why do you think this is a hardware error? Have you seen it before? ��
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Old 06-11-2016, 04:21 AM   #4
kmcarr
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Hi kmcarr,
Why do you think this is a hardware error? Have you seen it before? ��
Soleulloa,

It really couldn't be more clear; the error message itself told you that something went wrong with a piece of hardware, the Z motor, on your instrument. If your MiSeq stops mid run and throws up an error message the first thing to do is call Illumina Tech Support. They're the ones that can diagnose and fix the problem.
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Old 06-13-2016, 05:48 AM   #5
soleulloa
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Pleeeease.. any of you have seen that error before?
I'm afraid to run again and lose the reagents!
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Old 06-13-2016, 06:02 AM   #6
GenoMax
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@soleulloa: Any time there is an error (no matter what kind) you should contact Illumina tech support and have them take a look at the run remotely (as @kmcarr has said before). The error message does sound hardware related so if there is a problem you would not want to run again until the issue is fixed.

We have seen similar errors before but the problem is the cause can be very specific and can only be diagnosed by Illumina.
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Old 06-13-2016, 07:48 AM   #7
thermophile
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Don't run it again, call tech support. A field scientist or engineer will come out to check your z motor. They will replace the kit that was wasted by the error as long as you have a service contract.
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Old 06-13-2016, 09:34 AM   #8
Jessica_L
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To reiterate what everyone else is saying, you should definitely contact Illumina.

There is a chance that Z-motor issues can be related to trouble finding the best focal plane for imaging, i.e. something run/sample related rather than hardware, but those error messages usually look a bit different: "Best focus is too near edge of range: Autofocus would have moved Z to NaN, outside soft limits of -0.295001144426642 -0.00489885557335775: Get Move Z Motor To Focus Command", or something to that degree.

At what point in the run did it run stop? If you got far enough along to actually get cluster counts and the error message showed up mid-run, that's better evidence to support this being a hardware issue, as issues with focusing usually appear in cycle 1.
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Old 06-13-2016, 12:43 PM   #9
soleulloa
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Hi all,

I've contacted Illumina support immediatly. They ask for me some archives (sample sheet, runinfo, runparameter and InterOp) and finally the answer was:
YOUR SAMPLES ARE VERY CONCENTRATED AND THERE IS A FOCUS PROBLEM BECAUSE DENSITY CLUSTER IS SO HIGH

That answer is not satisfactory for me because Density Cluster for this run was 1266 K/mm2 and I've got similar values for other runs and all it is perfect.

I think that Illumina is not analyzing the problem in a right way.

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Old 06-13-2016, 12:46 PM   #10
thermophile
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call/email your FAS. Most at the general tech support are good but there are a few that aren't.

As far as clustering too dense, have you looked at thumbnails for all 4 channels? Do they look overclustered?
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Old 06-13-2016, 02:24 PM   #11
kmcarr
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Quote:
Originally Posted by soleulloa View Post
Hi all,

I've contacted Illumina support immediatly. They ask for me some archives (sample sheet, runinfo, runparameter and InterOp) and finally the answer was:
YOUR SAMPLES ARE VERY CONCENTRATED AND THERE IS A FOCUS PROBLEM BECAUSE DENSITY CLUSTER IS SO HIGH

That answer is not satisfactory for me because Density Cluster for this run was 1266 K/mm2 and I've got similar values for other runs and all it is perfect.

I think that Illumina is not analyzing the problem in a right way.

If a flow cell is over clustered you can not believe the cluster density number which the software states. If the clusters are packed so closely together the image analysis software can't separate them so the cluster density reported is way, way lower than the true number. Illumina's analysis may be entirely reasonable. Can you post some Thumbnails for this run please. Without those we would just be blindly guessing.
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