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  • #16
    We also have problem with LOT 93866020. It is better then the one Roche replaced, but it is not consistent. Some kits works as old good LOTs, when we have to wash 6-7 times to get rid off null beads, with some kits we have good sequencing data after about 10 washes, but some kits require 12, 15 or more washes with very low number of detected wells.
    I called today techsupport. They told that nobody complained about this LOT. Does anybody have a similar experience with LOT 93866020 as we do?

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    • #17
      Hey guys, I am joining this party late but I am also having trouble with Enrichment Kit Lot # 93866020. My enrichment rate for both samples I worked on using that lot number ended up being 2.5% to 3% enrichment. This has never happened before for me. I think this lot number is iffy but Roche says this kit is perfectly fine. Please beware of abnormally low enrichment values using this lot number. Please input any suggestions.

      I have contacted my FAS regarding this issue but I did not really receive an adequate answer to be honest. If they told you that no one complained about that lot number then they must be either mistaken or something else because I certainly did complain.
      Last edited by Aniki; 04-20-2012, 07:21 AM.

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      • #18
        Originally posted by vlee2 View Post
        We also have problem with LOT 93866020. It is better then the one Roche replaced, but it is not consistent. Some kits works as old good LOTs, when we have to wash 6-7 times to get rid off null beads, with some kits we have good sequencing data after about 10 washes, but some kits require 12, 15 or more washes with very low number of detected wells.
        I called today techsupport. They told that nobody complained about this LOT. Does anybody have a similar experience with LOT 93866020 as we do?
        As I mentioned on my earlier post I had the problem you described for this lot. I submitted the case yesterday. I was told by Roche people that I should not realy on forum info. However if was not for this forum I would not know a few lots of bead recovery reagents had problem. As a matter of fact I was the one who communicated to our FAS about this matter, to which he checked, confirmed the problem and got our lots replaced. However replacement lot does seem to have a problem as well.

        For lot 9386020 I was told "roughly 10 reports worldwide seem evident to date, among hundreds to thousands of other runs using the exact same lot without any problem. According to direct communication with responsible colleagues, currently this lot is being tracked." So I am waiting on their answer about my run with very low number of raw wells (about 25%).

        Today we did another enrichment using this same lot and everything worked normally. We will sequence next week. It might be another lot with good boxes and bad ones :S


        Roche should really come forward and release an official statement about the quality issues that have been found across several lots of the same reagent...

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        • #19
          We did another run with beads enriched using lot 9386020. We got over 1,6 million reads even though we had a problem while assembling the run: the tech forgot to incubate the beads with DBIM, she realized the mistake after the 2nd layer was already loaded. So we decided to recover the supernatant, add polymerase and cofactor plus some BB2, apply on the plate, let sit on the bench for about 10 min, centrifuge, and follow the protocol from that point. We were pleased surprised with the outcome.

          We still don´t have the official answer about our bad run. To me it looks like there is inconsistence on that lot as vlee2 mentioned.

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          • #20
            Originally posted by erikt View Post
            AJ, if you do sequence those beads I'd definitely be curious to know the results.
            Sorry I didn't respond to this earlier.

            I did sequence those beads, and the results were normal. Nothing spectacular, but just fine.

            I cleaned up some more emulsions yesterday using the same lot (93866020) and those enriched perfectly normal. I only needed to wash about 3 times, and I ended up with 6.5-8% enrichment.

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            • #21
              Our FAS told me today that lot 93866020 is under quarantine. It seems obvious some boxes are fine but some have problems. The run we did usinf beads recovered with this lot and that showed low number of raw wells is gonna be replaced.

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              • #22
                Originally posted by MissDNA View Post
                Our FAS told me today that lot 93866020 is under quarantine. It seems obvious some boxes are fine but some have problems. The run we did usinf beads recovered with this lot and that showed low number of raw wells is gonna be replaced.
                Not here. Info from FAS:

                "At this time lot #93866020, bead recovery buffer box, has not been quarantined within United States Roche nor Global Roche. There are complaints from U.S. customers that all the beads are attracted to the magnet during enrichment, but the report rate is currently lower than needed to quarantine the lot."

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                • #23
                  You are right, HMorrison. Our FAS had given me the info I posted over the phone but he wrote me yesterday saying:

                  "All lots of Bead Recovery Reagent will be evaluated, and if some lot presents more than 5% failure it will be collected. Until the present date, we received only 22 complaints for Lot # 93866020, which had 1000 units produced, including those reported in Brazil. This means that we have 2.2% of the lot with different complaints. This still does not represent a problem in the lot.

                  You should proceed with the use of lots 93866020 and 93848420, the latter, only units which have a normal pellet (in stock). Both lots have less than 5% of complaints. If you have problems, we will proceed with opening a case and replace it, according to the guidelines of our support. As to Lot 93852420, the recommendation is to not use."

                  So that is what we are going to do, as we have no other lots in stock in the lab nor in Brazil atm.

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                  • #24
                    Originally posted by MissDNA View Post
                    You are right, HMorrison. Our FAS had given me the info I posted over the phone but he wrote me yesterday saying:

                    "All lots of Bead Recovery Reagent will be evaluated, and if some lot presents more than 5% failure it will be collected. Until the present date, we received only 22 complaints for Lot # 93866020, which had 1000 units produced, including those reported in Brazil. This means that we have 2.2% of the lot with different complaints. This still does not represent a problem in the lot.

                    You should proceed with the use of lots 93866020 and 93848420, the latter, only units which have a normal pellet (in stock). Both lots have less than 5% of complaints. If you have problems, we will proceed with opening a case and replace it, according to the guidelines of our support. As to Lot 93852420, the recommendation is to not use."

                    So that is what we are going to do, as we have no other lots in stock in the lab nor in Brazil atm.
                    I do not have any other lots besides lot 93866020 as well. I have no choice but to use it. However, the last time I used this lot I achieved 2-3% enrichment rate and a raw well number of 25% or less. If I get another bad run, I will have to stop using this lot.

                    Comment


                    • #25
                      Originally posted by HMorrison View Post
                      Not here. Info from FAS:

                      "At this time lot #93866020, bead recovery buffer box, has not been quarantined within United States Roche nor Global Roche. There are complaints from U.S. customers that all the beads are attracted to the magnet during enrichment, but the report rate is currently lower than needed to quarantine the lot."
                      I currently just finished recoverying/enriching a different library. It turns out just as aweful as my previous attempt.

                      Recovery rate was good at 83%
                      Enrichment rate was abyssmal at 2.5%

                      Furthermore, at the melt step to separate the enrichment beads from DNA beads, the DNA beads were sticking to the enrichment beads on the outside quite extensively. I took pictures if anyone is interesed. I used lot 93866020 again. I will no longer attempt more until my FAS can assure me otherwise.

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