Team:Heidelberg/Templates/Del week11 AE

From 2013.igem.org

(Difference between revisions)
(Created page with "==08-07-2013== ===Amplification from FS_02 to FS_03; 5.3 kb=== [[File:20130708 DelAE DelBB DelG.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.0...")
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==08-07-2013==
==08-07-2013==
===Amplification from FS_02 to FS_03; 5.3 kb===
===Amplification from FS_02 to FS_03; 5.3 kb===
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[[File:20130708 DelAE DelBB DelG.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kb), lanes5-10=pSB4K5 (4.2kb), lanes11-13:DelG (6.4kb);
+
[[File:Heidelberg_20130708 DelAE DelBB DelG.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kb), lanes5-10=pSB4K5 (4.2kb), lanes11-13:DelG (6.4kb);
run at 100 V, 0.8 % gel (TAE)]]
run at 100 V, 0.8 % gel (TAE)]]
:'''Reaction'''
:'''Reaction'''
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! what !! µL   
! what !! µL   
|-
|-
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| ''D. acidovorans'' || 1
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| ''D. acidovorans'' DSM-39|| 1
|-
|-
| FS_02:  (1/10) || 2.5
| FS_02:  (1/10) || 2.5
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[[File:20130708 DelAE DelBB DelG cut.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), cut]]
+
[[File:Heidelberg_20130708 DelAE DelBB DelG cut.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), cut]]
:'''Conditions'''
:'''Conditions'''
{| class="wikitable"
{| class="wikitable"

Revision as of 01:30, 30 September 2013

08-07-2013

Amplification from FS_02 to FS_03; 5.3 kb

PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kb), lanes5-10=pSB4K5 (4.2kb), lanes11-13:DelG (6.4kb); run at 100 V, 0.8 % gel (TAE)
Reaction
what µL
D. acidovorans DSM-39 1
FS_02: (1/10) 2.5
FS_03: (1/10) 2.5
Phusion Master Mix 25
DMSO 2.5
dd H2O 19


PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), 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 of DelAE worked
  • bands were cut out and DNA purified using QIAquick Gel Extraction Kit