Team:Heidelberg/Templates/Del week11 AE

From 2013.igem.org

(Difference between revisions)

Latest revision as of 21:38, 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

@@ Line 1: / Line 1: @@
 ==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 (08.07) 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)]]
 :'''Reaction'''
@@ Line 8: / Line 9: @@
 ! what !! µL
 |-
-| ''D. acidovorans'' || 1
+| ''D. acidovorans'' DSM-39|| 1
 |-
 | FS_02:  (1/10) || 2.5
@@ Line 22: / Line 23: @@
-[[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'''
 {| class="wikitable"