[Aniki]

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.

[Aniki]

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.

[MissDNA]

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.

[HMorrison]

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."

[MissDNA]

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.

[ajthomas]

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.

[MissDNA]

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.

[MissDNA]

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...

[Aniki]

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.

[vlee2]

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?

[MissDNA]

FAS told me the lot was not flagged. We did the sequencing run and what was noticiable was a very low raw well number, which made me remember a post I saw on this forum:

"93852420 we had problems with low % recovery (below 5%). If we used a different kit with the same cpb, the emulsion would work every time. I was informed that the problem with this lot number though does not cause emPCR failure but, and I quote, "Rather than observing emPCR failure, this particular lot number was associated with a low raw well number in analysed seq runs"."

Once again I will contact support! This has become a daily routine for me. So many problems!

[MissDNA]

We did a LV recovery today using lot 93866020 for the first time. It was necessary 12 washes while normally we do around 6-7. At the end we got around 2.5 million beads per cup, which is perfect for sequencing. Sequencing will happen tomorrow. We will be using XLR70. I am looking forward the results and certainly will check the filters.

Still waiting on our FAS response to see if this lot has a flag or not.

[erikt]

Unfortunately my results today were somewhat inconclusive. 1 cpb yielded 2.5%, and 3 cpb 2.9%. However, the first wash did remove the largest portion of the beads, and far fewer washes were necessary, so that is encouraging. I received my replacement kits today; those are also lot 93866020, so I have little choice but to carry on. I'll follow up if I run into any trouble with this lot.

[MissDNA]

Roche is going to replace all our 93848420 and 93852420 boxes. They said we should discard all the 8420 boxes but we should save the boxes from 2420 that present normal quantity of beads to be used in case of need.

What we have been getting so far is 93866020, which I hope offers no problems.

We still have a few boxes of 93837120, which is also listed here but Roche says is fine.

[erikt]

I'll post the results.

AJ, if you do sequence those beads I'd definitely be curious to know the results.