09/09/13

From 2013.igem.org

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==Measurement of DNA concentration==
==Measurement of DNA concentration==
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{| border=1
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|Samples ||Volume (ul)||Conc (ng/ul)||280/260||260/280
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|-
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|TOD X1||31.6||86.5||1.87||1.92
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|-
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|TOD X2||31.7||58.7||1.89||1.26
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|-
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|TOD X3||32.7||56.6||1.91||1.88
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|-
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|TOD X4||31.6||55.8||1.88||1.65
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|-
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|TOD X5||31.7||44.4||1.87||1.78
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|-
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|TOD X6||31.7||70.5||1.88||1.88
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|}
==Double Digestion of pSB1C3/TOD genes==
==Double Digestion of pSB1C3/TOD genes==

Revision as of 16:27, 9 September 2013

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Miniprep of overnight broth

  • Following on from 06/09/13 result,overnight broth was prepared yesterday from the single colonies that had grown on each of the 2 plates shown on 06/09/13.
  • The broth, which contains cells that have been transformed with pSB1C3/TOD genes, was minipreped using the Omega Bio-Tek kit.
  • This would enable us to confirm which cells have the recircularised plasmid and those with the ligated product.

Measurement of DNA concentration

Samples Volume (ul)Conc (ng/ul)280/260260/280
TOD X131.686.51.871.92
TOD X231.758.71.891.26
TOD X332.756.61.911.88
TOD X431.655.81.881.65
TOD X531.744.41.871.78
TOD X631.770.51.881.88

Double Digestion of pSB1C3/TOD genes

  • The pSB1C3/TOD genes were double digested with (XbaI and SpeI) & (EcoRI and PstI)
  • The master mix for the (XbaI and SpeI) was;

60ul of cut smart buffer 5ul of XbaI 10ul of SpeI 273ul of 5M Tris

  • The master mix for the (EcoRI and PstI) was;

60ul of cut smart buffer 5ul of XbaI 5ul of SpeI 278ul of 5M Tris

All 36 tubes were incubated overnight in 37 degrees room.

  • Tomorrow the plasmids that were isolated from the miniprep would be run on a 1% agarose gel to distinguish between cells that have the pSB1C3/TOD genes from those that have the recircularised pSB1C3 vector.