MiSeq flow rate won't measure

[brc-seq]

Hello,

I am having an issue with my MiSeq and was wondering if any of you have had a similar issue or any ideas of what could be causing the issue.

I went to start a run last friday night and the flow rate failed to acquire for the final check. The error said it could not detect a flow rate. I tried pumping PR2 without any success. And then did a line wash after which the instrument said there may be a clog contact illumina support.

I was advised to try different flow cells, using the wash flow cell the same errors occurred. To do a power cycle and multiple washes (after multiple line washes and maintenance washes there still has been no success). Also, I have done the Volume Test which passed and have done multiple power cycles.

So if the flow cell isn't the issue and If the volume test pass indicates that it isn't a pump, valve, or sensor, then is it certainly a clog? If so how much more washing can be done before we pay for a technician to come out?

One last odd thing is there is an "@@@---failed to get temperature" warning that keeps popping up and has done so through multiple power cycles. Illumina says this wouldn't affect anything and also is not related to the issue, but I thought I would include it since its odd it started happening at the same time.

I'm not sure I understand the issue completely, and I am out of ideas for testing the instrument. If you have any ideas to share it would be much appreciated.

Ryan V
Biomedical Research Centre
Vancouver, BC

[GenoMax]

I suppose you don't have a maintenance contract on this machine and are trying to determine if you need to ask a technician to come out and service the machine? Probably not what you want to hear but it sounds like you do.

[brc-seq]

GenoMax, you have hit the nail right on the head. We are starting to plan for a technician to come out, but if there are any other ideas I am still open to try.

Thanks for confirming our fears!

[GenoMax]

Consider a service contract in future, if you use the machine on a regular basis. Think of it as part of the cost of using the latest technology.

Good luck with the repair.

[brc-seq]

We let it lapse as we are switching our work onto a NextSeq which should be arriving in the next few weeks. Now we will be torn into deciding if it is worth fixing the miseq as our plans were to use it only for the occasional amplicon seq. Anyhow, as you say, these are all the costs of using the newest technologies and at least our NextSeq should be arriving shortly.

[GenoMax]

You may want to keep that MiSeq going till you convince yourself that the NextSeq 500 is a solid replacement for your needs. See this thread: http://seqanswers.com/forums/showthread.php?t=40741

[brc-seq]

Always a good idea, but the NextSeq seems to fit our needs well and the shortcomings aren't to big a concern for us. We are focusing more on throughput and expression, but i guess we will see. You are right though, the MiSeq may still hold more value than we expect. Point to getting the engineer in.

Cheers.

[cmbetts]

I'm not sure how helpful this will be, since if you have the same problem I had, the solution required an Illumina repair technician, but here goes.

My former company's MiSeq went through a phase where it would randomly mangle the gaskets going into/out of the flowcell, which caused runs to fail in a similar fashion. After having Illumina support come out a few times, they finally got the problem under control.
A quick test to see if you are having the same problem would be to compare the gaskets on a failed vs successful flow cell (assuming you kept them). Our flowcells from failed runs had a very noticeable change in shape from circular to a sort of closed off oval.

[brc-seq]

cmbetts, thanks for the suggestion. However, I don't believe this to be a flow cell issue. We tried multiple flowcells and they all failed, also I found another MiSeq on campus which I was able to utilize my reagents for a run last night (this included the flowcell). Seems like there are many potential issues and I will be setting up for a Technician to come out.

Many thanks.

[d00b]

Miseq flowcell bubble and gasket

Miseq flowcell defective gasket causes premature running stop
[IMG][/IMG]

[HongjaePark]

Same problem here

Hi,

I wonder whether you finally solved your problem.

I have exactly the same problem (even "@@@---failed to get temperature" message) and in my case, technicians didn't do much help.

They keep saying there is no problem with flow rate but error almost always (may be 9 out of 10 cases?) pops out.

The only way I solve this problem is keep washing and turn on and off until it works..

Please let me know if you solve it. I would do the same.

[d00b]

do you have kakao id? i'd like to call you. thanks

[Brian Bushnell]

Miseq flowcell defective gasket causes premature running stop
[IMG][/IMG]

d00b,

Thanks for the excellent post. That is exactly what I would hope for were that my problem.

[HongjaePark]

Flow rate issue

Hi d00b,

My Kakao id is 'sntiger735'.

Before calling, I want to tell you my issue isn't about premature stop.

I have errors at initial step and once I get out of it, it worked fine until end.

Also, I checked holes of the flow cell and it looks fine!

Can you still be my savior? ^^ thanks.

[d00b]

To Hongjae

I see. then your issue is very simple. Just clean up your flow cell and check the gasket surface. you also need to clean the injector nozzle part (orange color) and flow cell holder before you insert flow cell.

if you still have error, request new flowcell to illumina. They will replace the flow cell.