Team:Heidelberg/Templates/Del week10 AE

From 2013.igem.org

(Difference between revisions)
Line 8: Line 8:
! what !! µL   
! what !! µL   
|-
|-
-
| ''D. acidovorans'' || 1
+
| ''D. acidovorans'' DSM-39 || 1
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| FS_02:  (1/10) || 1
| FS_02:  (1/10) || 1

Revision as of 01:29, 30 September 2013

06-07-2013

Amplification from FS_02 to FS_03; 5.3 kb

PCR for amplification of del fragments E-G and A-E, AE+=PCR AE with DMSO, AE-=PCR without DMSO, M= Marker; wanted amplicon sizes are: AE=5.3kb, EG=11.0
Reaction
what µL
D. acidovorans DSM-39 1
FS_02: (1/10) 1
FS_03: (1/10) 1
Phusion Master Mix 10
DMSO 1/-
dd H2O 6/7
PCR for amplification of del fragments AE and EG, cut
Conditions
Biorad MyCycler
Cycles-PCR temperature [°C] Time [s]
1 98 5
12 98 1
66 ↓ 0.5 5
72 2:30 min
18 98 1
63 5
72 2:30 min
1 72 10 min
1 4 inf

Results:

  • Amplification worked with 5% DMSO
  • Repeat Amplification with the same protocol to increase concentration when DNA is extracted from gel slices
Retrieved from "http://2013.igem.org/Team:Heidelberg/Templates/Del_week10_AE"