Pipetting, contamination....

ubwasp66 via methods%40net.bio.net (by ubwasp66 from peoplepc.com)
Sat Feb 17 22:17:49 EST 2007


   Suggestions below.
   Good luck.
   Teresa<= br>

     -----Original Message-----
     From: Gaelle ayyildiz
     Sent: Feb 17, 2007 6:33 PM
     To: ubwasp66
     Subject: Re: Pipetting, contamination....
     Hey Teresa,
     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,
     sterile= tube;
     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
     them.
     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
     tube?
     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
     reactions.
     N= ot sure if this answers your question but hope that helps,
     Teres= a
     -----Original Message-----
     >From: Gaelle ayyildiz
     >Sent: Feb 16, 2007 3:42 AM
     >To: methods= @magpie.bio.indiana.edu
     >Subject: Pipetting, contamination....
     >= ;
     >Hi all,
     >
     > 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..
     >
     >
     >Gaelle
     >
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     Gaelle

      
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