Bead Enrichment - infinite washes

[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.

[HMorrison]

Originally Posted by 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.

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]

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.

[Aniki]

Originally Posted by 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.

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.

[Aniki]

Originally Posted by HMorrison

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.