Team:Evry/Interlab Study/08-22-2014

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<h2>Contructions PSB1C3 with I20260, J23101-E1010 and K823012-E1010</h2>
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<br/> Transformation plate observation:
<br/> Transformation plate observation:
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<br/>- BBa_J23101: nothing grown
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<br/>- BBa_J23101: 11 isolated colonies
<br/>- BBa_K823012: 100-150 isolated colonies
<br/>- BBa_K823012: 100-150 isolated colonies
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<br/> A PCR was performed on 8 colonies for BBa_K823012 following the protocol Table 3 and 4.
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<br/> A PCR was performed with 8 colonies for BBa_K823012 and for BBa_J23101 following the protocol Table 3 and 4.
<br/> To verify PCR products, 10 µl was loaded on a 1% agarose gel with 2 µl of loading dye 6X.
<br/> To verify PCR products, 10 µl was loaded on a 1% agarose gel with 2 µl of loading dye 6X.
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     <center>1% agarose gel of PCR products of the eight selected colonies</center>
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     <center>1% agarose gel of PCR products. Lane 1 to 8: BBa_K823012 PCR products and Lane 9: Purple 2-Log. ladder NEB</center>
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<br/> We expected to obtain a band between 300 and 400 bp.
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<br/> We expected to obtain a band between 300 and 400 bp. So we decided to amplify the colony n°8. The colony was seed in 5 ml of LB medium and 5 µl of chloramphenicol. Culture was incubated overnight at 37°C with 200 rpm agitation.
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    <center>1% agarose gel of PCR products. Lane 10 to 17: BBa_J23101 PCR products and Lane 9: Purple 2-Log ladder NEB</center>
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<br/> We expected to obtain a band around 1200 bp. So we decided to amplify the colony n°2 corresponding to lane 11. The colony was seed in 5 ml of LB medium and 5 µl of chloramphenicol. Culture was incubated overnight at 37°C with 200 rpm agitation.
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Latest revision as of 17:59, 19 September 2014

Picture

Contructions PSB1C3 with I20260, J23101-E1010 and K823012-E1010


Transformation plate observation:
- BBa_J23101: 11 isolated colonies
- BBa_K823012: 100-150 isolated colonies

A PCR was performed with 8 colonies for BBa_K823012 and for BBa_J23101 following the protocol Table 3 and 4.
To verify PCR products, 10 µl was loaded on a 1% agarose gel with 2 µl of loading dye 6X.

IMAGE
Symbol
1% agarose gel of PCR products. Lane 1 to 8: BBa_K823012 PCR products and Lane 9: Purple 2-Log. ladder NEB

We expected to obtain a band between 300 and 400 bp. So we decided to amplify the colony n°8. The colony was seed in 5 ml of LB medium and 5 µl of chloramphenicol. Culture was incubated overnight at 37°C with 200 rpm agitation.
IMAGE
Symbol
1% agarose gel of PCR products. Lane 10 to 17: BBa_J23101 PCR products and Lane 9: Purple 2-Log ladder NEB


We expected to obtain a band around 1200 bp. So we decided to amplify the colony n°2 corresponding to lane 11. The colony was seed in 5 ml of LB medium and 5 µl of chloramphenicol. Culture was incubated overnight at 37°C with 200 rpm agitation.

Aug 22

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