Team:Paris/August 4

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Latest revision as of 15:22, 6 August 2008

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PCR Screening of Ligation Transformants

Use of 8 clones of Ligation transformants n°L100 for screening PCR

Protocol of screening PCR

Mix

Name	Vol (µl)	Concentration
Quick Load	25µl	2X
OligoF_VF2 (O18)	1µl	10µM
OligoR_VR (O19)	1µl	10µM
water	23µl

50µl of Mix PCR by tube/clone
one toothpick of each clone's colony by tube
Program : Annealing 55°C - Time élongation 1'30" - Number cycle : 29

Conditions of electrophoresis

10µl of ladder 1 kb
20µl of screening PCR
migration ~30min at 100W on 0,8% gel

Results for L100

L100= D110 + D130 = RBS-tetR-ECFP-Ter

Band	1-->8
Expected size	1042 pb
Measured size	1100 pb
Gel : L100(1-8)

==> Conclusion : we obtain a low quantity of PCR products, but at the good size.
So now, we know that we can amplify the clones that we want for L100 transformants bacteria.

Revision as of 15:21, 6 August 2008 (view source) Fanny.c (Talk \| contribs) (→Results for L100) ← Older edit		Latest revision as of 15:22, 6 August 2008 (view source) Fanny.c (Talk \| contribs) (→Results for L100)
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	==> '''Conclusion :''' we obtain a low quantity of PCR products, but at the good size.		==> '''Conclusion :''' we obtain a low quantity of PCR products, but at the good size.
-	So now, we know that we can amplify ~~one of~~ the clones of L100 transformants bacteria.	+	<br>So now, we know that we can amplify the clones that we want for L100 transformants bacteria.