(by ubwasp66 from peoplepc.com)
Sat Feb 17 22:17:49 EST 2007
From: Gaelle ayyildiz
Sent: Feb 17, 2007 6:33 PM
Subject: Re: Pipetting, contamination....
First of all, thanx a million for replying m= y email. u've given
me a useful answer. Actually, i still have a question = about the
way of pipetting from a wide-mouth bottle where anything in the "body" of the pipette may cause contamination. what should i do?
A:= You should use sterile pipettes, if going back to the same
solution in the= "wide-mouth bottle" more than once take an aliquot
that will cover your reaction requirements and place in a small,
and, moreover, how can i handle the cover of the 1.5ml-pr esterilized tubes in a way that guaranties the sterility?
A: If keep= ing "sterile working conditions" is a must, work in a
biosafety cabinet (= =hood). Take a subset of tubes by dropping
them on the top of a presteril= ized container (glass petri dish
for example) and take each tube by holding= to the bottom, never
the lid. If you want to, sterilize a set of pliers an= d work with
have u worked with PCR before? i've a problem = in pipetting, i
always have a drop remains in the tip & doesn't go int= o the PCR
A;Always work with calibrated pipets. The "leftover" dro= p does
not pose a problem to your reaction (usually make a 10% extra of
you= r PCR mix to account for pippeting errors).
If dealing with dense, oil-l= ike chemicals, look for special tips
for that purpose.
im re= ally grateful & really sorry for interrupting u...
ubwas= p66 <ubwasp66 from peoplepc.com> wrote:
Gaelle, You should use barrier-like tips (filter tips) (check the Gilson
we= bsite for specs).
Also, you should have a dedicated set of pipe= ttes and tip racks
(preferably located in separate room from your "nucleic acid" room
or ins= ide a dedicated "clean" PCR workstation) to set up PCR
N= ot sure if this answers your question but hope that helps,
>From: Gaelle ayyildiz
>Sent: Feb 16, 2007 3:42 AM
>To: methods= @magpie.bio.indiana.edu
>Subject: Pipetting, contamination....
> would u please tell me how i can pipet= te my regents without
contaminating them with the micropipette (i.e. Gilso= n pipette)?
> i would really appreciate if u can show me = pictures or figures
of the safe way of pipetting.
> Thanx= a million..
>We won't tell. Get more on shows you hate to love<= br>>(and love
to hate): Yahoo! TV's Guilty Pleasures list.
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