Team:TU Darmstadt/Labjournal/Transport

From 2012.igem.org

(Difference between revisions)
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* First [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Colony_PCR colony PCR] of ''Comamonas testosteroni'' for isolation of the following genes: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808004 tctA 505aa] (1), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808005 tctB 197aa] (2), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808002 tctA_503aa] (3), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808003 tctB 162aa] (4), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808001 tphC 322aa] (5), 50µl reaction mixture each; analysis of [http://en.wikipedia.org/wiki/Polymerase_chain_reaction PCR] products by [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Agarose_gel_electrohporesis agarose gel electrohporesis]
* First [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Colony_PCR colony PCR] of ''Comamonas testosteroni'' for isolation of the following genes: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808004 tctA 505aa] (1), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808005 tctB 197aa] (2), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808002 tctA_503aa] (3), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808003 tctB 162aa] (4), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808001 tphC 322aa] (5), 50µl reaction mixture each; analysis of [http://en.wikipedia.org/wiki/Polymerase_chain_reaction PCR] products by [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Agarose_gel_electrohporesis agarose gel electrohporesis]
:https://static.igem.org/mediawiki/2012/1/15/W1_1_kolo_pcr.png
:https://static.igem.org/mediawiki/2012/1/15/W1_1_kolo_pcr.png
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* PCR product purification using the Promega-Kit
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* Nanodrop measurements of the purified PCR products:
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:(1) : 19.5 ng/µl 260/280=1.58
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:(2) : 61.9 ng/µl 260/280=1.75
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:(3) : 75.1 ng/µl 260/280=1.75
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:(4) : 93.3 ng/µl 260/280=1.85
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:(5) : 91.3 ng/µl 260/280=1.58
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* [ https://2012.igem.org/Team:TU_Darmstadt/Protocols/Restriction_digest Digestion] of PCR products by the restriction enzymes SpeI and EcoRI, heat inactivation after digestion
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* Analysis of restriction digest by [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Agarose_gel_electrohporesis Agarose gel electrohporesis]
 +
:https://static.igem.org/mediawiki/2012/1/15/w1_2_restriktionsverdau_s8.png
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* Purification of the bands gained form gel electrophoresis (Promega-Kit)
 +
* Nanodrop measurements of the purified products:
 +
:(1) : 8.3 ng/µl 260/280=1.94
 +
:(2) : 8.9 ng/µl 260/280=1.80
 +
:(3) : 13.0 ng/µl 260/280=1.57
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:(4) : 1.5 ng/µl 260/280=3.08
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:(5) : 3.7 ng/µl 260/280=2.26
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===week 2 (28.05.-01.06.12)===
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* Purified products from 1. week were [https://2012.igem.org/Team:TU_Darmstadt/Protocols/DNA_Ligation ligated] into pSB1A2 and subsequently [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Bacterial_transformation transformed] into DH5α (in spite of low concentration)

Revision as of 21:04, 8 September 2012

Transport

Week 1 (21.-25.05.12)

  • First colony PCR of Comamonas testosteroni for isolation of the following genes: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808004 tctA 505aa] (1), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808005 tctB 197aa] (2), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808002 tctA_503aa] (3), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808003 tctB 162aa] (4), [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808001 tphC 322aa] (5), 50µl reaction mixture each; analysis of [http://en.wikipedia.org/wiki/Polymerase_chain_reaction PCR] products by agarose gel electrohporesis
W1_1_kolo_pcr.png
  • PCR product purification using the Promega-Kit
  • Nanodrop measurements of the purified PCR products:
(1) : 19.5 ng/µl 260/280=1.58
(2) : 61.9 ng/µl 260/280=1.75
(3) : 75.1 ng/µl 260/280=1.75
(4) : 93.3 ng/µl 260/280=1.85
(5) : 91.3 ng/µl 260/280=1.58
w1_2_restriktionsverdau_s8.png
  • Purification of the bands gained form gel electrophoresis (Promega-Kit)
  • Nanodrop measurements of the purified products:
(1) : 8.3 ng/µl 260/280=1.94
(2) : 8.9 ng/µl 260/280=1.80
(3) : 13.0 ng/µl 260/280=1.57
(4) : 1.5 ng/µl 260/280=3.08
(5) : 3.7 ng/µl 260/280=2.26

week 2 (28.05.-01.06.12)

  • Purified products from 1. week were ligated into pSB1A2 and subsequently transformed into DH5α (in spite of low concentration)
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