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November 2018

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From:
dev mistry <[log in to unmask]>
Reply To:
The Pharmaceutical Microbiology Forum Email List <[log in to unmask]>
Date:
Sat, 10 Nov 2018 00:48:21 +0000
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Hello,
I would let the new analyst practice with water addition to an empty plate before they start performing the real testing. Pipetting training could also help with better pipetting technique. 

Lonza reagents has to be pre-incubated for 10 minutes. Not that that would cause any problems with reaction times but check temperature uniformity test to see if there is any issue with uniform plate heating. Sometime different reagent lots can make difference as well.
Single channel is not appropriate for lysating.
Regarding using multi channel pipette or repeater pipette , you can use any as long as you are quick enough to finish the whole plate within ~2 minutes. For new analysts, I would recommend using repeater pipettes. Eppendorf has new electronic repeater pipette  (E3 and E3X)that can help with faster plating- of course that will also require practice to become familiar with the pipette.
Also, always start platting left to right and top to bottom. Ask reagent manufacturer to see if they have any recommendation for additions of lysate in a particular way. 

Regards,
DeX

Sent from my iPhone

> On Nov 9, 2018, at 6:19 PM, Sattelberg, Nicholas <[log in to unmask]> wrote:
> 
> I'm curious how others are adding lysate to their endotoxin plates before loading for kinetic methods.
> 
> Are you:
> 
> 
>  1.  Using a Multichannel pipette and reagent reservoir
>  2.  Using a repeat pipettor
>  3.  Using a single channel pipette
> 
> Obviously you want to add lysate as quickly as possible since the reaction starts as soon as it's added to a pre-warmed plate. I have seen plenty of plates that had funky results because new techs in training were too slow with this last step. There's recently been a debate as to which method is best. I've heard claims that multichannels should never be used because there's a risk of contamination using the reagent reservoir. I'm not sure I buy that argument if you're properly changing tips each time and practicing aseptic technique.
> 
> Nick Sattelberg
> Lead Scientist, Microbiology
> 
> 
> 
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Please take a second to visit our sponsors' web sites and say thank you for their support of this service.
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Science Advisory Board https://www.scienceboard.net/

Sartorius https://promotions.sartorius.com/real-heroes

Steris - http://www.sterislifesciences.com/

Avista Pharma Solutions  http://www.avistapharma.com

MilliporeSigma  http://www.emdmillipore.com/

Charles River Laboratories - http://www.criver.com/

Veltek Associates, Inc - http://www.sterile.com

Microbiologics, Inc. - http://www.microbiologics.com

Nelson Labs - http://www.nelsonlabs.com

BD Diagnostic Systems - http://www.bd.com/ds/

Boston Analytical http://www.bostonanalytical.com/

Associates of Cape Cod, Inc. - http://www.acciusa.com/


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The nature of this service is to provide a medium for communication.  The specific statements and endorsements of individuals participating in the discussions are not necessarily those of The Microbiology Network, Inc., the PMF, or the sponsors of the list.

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