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  • pmiguel
    replied
    [QUOTE=gilly;18556]
    Originally posted by vlee2 View Post

    Hi we've seen this too. 750K for each region on a 2 region plate when we'd expect 1.1 million. We’ve re-counted the beads and I’m sure we’re loading the correct amount.
    Vlee et al.,

    I have delved into this issue previously:

    Pyrosequencing in picotiter plates, custom arrays for enrichment/decomplexing. (Roche)


    (Where I come to the wrong conclusion about the cause of the issue. We corrected our deviation from protocol, and the results did not change.)

    and a related thread about what does constitute a raw well:

    Pyrosequencing in picotiter plates, custom arrays for enrichment/decomplexing. (Roche)


    Basically the results we had been getting around that time, despite the lower numbers of raw wells, were decent, so we were not employing coercive interrogation methods on Roche representatives. Now the consensus seems to be: problem with multiple emPCR kits/batches.

    --
    Phillip

    Leave a comment:


  • pmiguel
    replied
    Originally posted by 454User View Post
    I just placed an order today for several SVemPCR kits and was informed that they are on backorder indefinitely.

    The XLR70 kits are supposed to ship as soon as the last component holding up the QC passes.
    My sale rep says the XLR70 issue was just the case of demand out stripping the batch size. She believed people might have heard "Roche back orders" and decided to stock up (on everything.)

    Big shipment is said to be going out today.

    Leave a comment:


  • LMcSeq
    replied
    We've had the same problem.

    Leave a comment:


  • gilly
    replied
    [QUOTE=vlee2;18550]
    Originally posted by pmiguel View Post
    Well, that is mysterious. I inferred that the problem was with the emPCR kits, because when I mentioned the "Low Number of Raw Wells" issue I was having at the User's Group meeting, the problem was immediately blamed on issues with emPCR kits.

    Hi Phillip,
    Could you tell more about "Low number of raw wells" issue. We have this issue almost every run lately and reported to Roche multiple times, but I hear from from you first time that this is not only our problem and the cause is emPCR kit.
    Thanks,
    Vlad
    Hi we've seen this too. 750K for each region on a 2 region plate when we'd expect 1.1 million. We’ve re-counted the beads and I’m sure we’re loading the correct amount.

    Leave a comment:


  • BaCh
    replied
    [QUOTE=vlee2;18550]
    Originally posted by pmiguel View Post
    Could you tell more about "Low number of raw wells" issue. We have this issue almost every run lately and reported to Roche multiple times, but I hear from from you first time that this is not only our problem and the cause is emPCR kit.
    The information politics of Roche/454 is ... improvable.

    Reminds me of the Titanium kits in the first half of last year which completely failed to give good data on GC rich (~ >= 62%) genomes or genome regions. Has never been made really public and I still meet people who paid quite some money and ask why they don't get good assemblies. The questions they ask and the symptoms they report make me immediately think that they must have sequenced it in that time frame. I'm right 90% of the time.

    Leave a comment:


  • vlee2
    replied
    [QUOTE=pmiguel;18390]Well, that is mysterious. I inferred that the problem was with the emPCR kits, because when I mentioned the "Low Number of Raw Wells" issue I was having at the User's Group meeting, the problem was immediately blamed on issues with emPCR kits.

    Hi Phillip,
    Could you tell more about "Low number of raw wells" issue. We have this issue almost every run lately and reported to Roche multiple times, but I hear from from you first time that this is not only our problem and the cause is emPCR kit.
    Thanks,
    Vlad

    Leave a comment:


  • 454User
    replied
    Originally posted by pmiguel View Post
    Anyone having trouble getting emPCR kits?
    I just placed an order today for several SVemPCR kits and was informed that they are on backorder indefinitely.

    The XLR70 kits are supposed to ship as soon as the last component holding up the QC passes.
    Last edited by 454User; 05-11-2010, 04:32 PM.

    Leave a comment:


  • pmiguel
    replied
    Originally posted by rbarry View Post
    They are telling me now that it's the XLR70 sequencing kits that are on backorder with no projected date as to when they will be available.
    Well, that is mysterious. I inferred that the problem was with the emPCR kits, because when I mentioned the "Low Number of Raw Wells" issue I was having at the User's Group meeting, the problem was immediately blamed on issues with emPCR kits. Maybe these are two separate issues then?

    Anyone having trouble getting emPCR kits?

    --
    Phillip

    Leave a comment:


  • rbarry
    replied
    My guess is that either this QC step is now a bottleneck or that so many lots of product from this vendor are being rejected that the supply of this vendor's product is bottlenecking the supply of emPCR kits.
    They are telling me now that it's the XLR70 sequencing kits that are on backorder with no projected date as to when they will be available.

    Leave a comment:


  • pmiguel
    replied
    Originally posted by rbarry View Post
    unfortunately all she will tell me is that she wishes she knew more
    Strange. As mentioned upthread, at the 2nd Users Group meeting last week, we were told that due to problems in QC from a "vendor", 454 users had seen variable results from a number emPCR kits from multiple lots. Because of this issue Roche had implemented additional QC that had (temporarily) apparently resulted in production delays.

    My guess is that either this QC step is now a bottleneck or that so many lots of product from this vendor are being rejected that the supply of this vendor's product is bottlenecking the supply of emPCR kits.

    --
    Phillip

    Leave a comment:


  • rbarry
    replied
    unfortunately all she will tell me is that she wishes she knew more

    Leave a comment:


  • nickloman
    replied
    We were sent a circular letter from Roche describing the problems. I'd re-post but they asked us not to. Get on the case of your account manager?

    Leave a comment:


  • LMcSeq
    replied
    At the user's group meeting they said that there is a large backorder on sequencing reagents. They have tightened up their quality requirements from vendors.

    Leave a comment:


  • rbarry
    started a topic 454 supply issues?

    454 supply issues?

    Does anyone know what's going on with all the supply interruptions at Roche/454? I can't seem to get a straight answer about anything from them.

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