Team:Heidelberg/Templates/Del week15 L

From 2013.igem.org

Contents

07-08-2013

Results Sequencing

Send the following samples (amplified fragments of D. acidovorans DSM-39 and backbone pSB4K5 from the partsregistry) to GATC for sequencing:

  • DelL (FS_14 to FS_15) with Primer FS_14
  • DelL (FS_14 to FS_15) with Primer FS_15

Alignment of DelL(FS_14 to FS_15) with Primer FS_14

Alignment of DelL(FS_14 to FS_15) with Primer FS_15

Amplification of DelL (FS_14 to FS_15; 1.4 kb)

Amplifcation of DelOP and DelL (07.08); run at 100 V, 0.8 % gel (TAE)
Amplifcation of DelOP and DelL (07.08), cut


Reaction
what µl
D. acidovorans SPH-1 1
FS_14: (1/10) 2.5
FS_15: (1/10) 2.5
Phusion Master Mix 25
DMSO 2.5
dd H2O 16.5


Conditions
Biorad MyCycler
Cycles-PCR temperature [°C] Time [s]
1 98 10
12 98 1
66 ↓ 0.5 5
72 42
18 98 1
63 5
72 42
1 72 12 min
1 4 inf

Results:

  • Amplification worked very well, we had bright bands on the right height.
  • Bands were cut out and DNA purified using QIAquick Gel Extration Kit.

08-08-2013

Concentration measurement of DelL

Fragment Primer Date PCR Concentration
DelL FS14-FS15 07-08-2013 51 ng/µl

09-08-2013

Amplification of DelL (FS_14 to FS_15; 1.4 kb)

Amplifcation of DelOP (I), DelFG and DelL ; run at 100 V, 0.8 % gel (TAE)(09.08)
Amplifcation of DelOP (I), DelFG and DelL ; run at 100 V, 0.8 % gel (TAE)(09.08)
Reaction
what µl
D. acidovorans 1
FS_14: (1/10) 2.5
FS_15: (1/10) 2.5
Phusion Master Mix 25
DMSO 2.5
dd H2O 16.5
Conditions
Biorad MyCycler*
Cycles-PCR temperature [°C] Time [s]
1 98 10
12 98 1
66 ↓ 0.5 5
72 42
18 98 1
63 5
72 42
1 72 12 min
1 10 inf

Results:

  • Amplification worked very well, we had bright bands on the right height.
  • Bands were cut out and DNA purified using QIAquick Gel Extration Kit.
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