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

From 2014.igem.org

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.

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.

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