Team:Heidelberg/Templates/Del week15 AE

From 2013.igem.org

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File:Heidelberg_AE-FS 02.txt | Sequencing of DelAE(FS_02 to FS_03) with Primer FS_02
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File:AlignementAE 02Sequenzierung.clustal.txt | Alignment of DelAE(FS_02 to FS_03) with Primer FS_02
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File:Heidelberg_AE-FS 03.txt | Sequencing of DelAE(FS_02 to FS_03) with Primer FS_03
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File:AlignementAE 03Sequenzierung.clustal.txt | Alignment of DelAE(FS_02 to FS_03) with Primer FS_03
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File:Heidelberg_AlignementAE 02Sequenzierung.txt | Alignement of expected sequence of DelAE and the sequencing results with primer FS_02
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File:Heidelberg_AlignementAE 03Sequenzierung.txt | Alignement of expected sequence of DelAE and the sequencing results with primer FS_03
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Latest revision as of 19:17, 4 October 2013

Contents

07-08-2013

Sequencing Results

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

  • AE (FS_02-FS_03) with Primer FS_02
  • AE (FS_02-FS_03) with Primer FS_03

Alignment of DelAE(FS_02 to FS_03) with Primer FS_02

Alignment of DelAE(FS_02 to FS_03) with Primer FS_03

08-08-2013

Amplification from FS_02 to FS_03; 5.3kb

Amplifcation of DelAE and DelEF and DelOP; run at 100 V, 0.8 % gel (TAE)(07.08)
Amplifcation of DelAE and DelEF and DelOP; run at 100 V, 0.8 % gel (TAE)(07.08)
Reaction
what µL
D. acidovorans SPH-1 1
FS_02: (1/10) 4
FS_03: (1/10) 4
Phusion flash Master Mix 10
Conditions
Biometra TProfessional Basic
Cycles temperature [°C] Time [s]
1 98 10
30 98 1
58 5
72 1:30
1 72 7min
1 12 inf

Results:

  • Amplification of DelAE was repeated successfully with the new strain SPH-1
  • bands were cut out and DNA purified using QIAquick Gel Extraction Kit
  • if fragment is used in Gibson assembly the amplification has to be repeated to increase the amount of product
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