Team:HokkaidoU Japan/Proof

Team:HokkaidoU Japan - 2016.igem.org

 

Team:HokkaidoU Japan

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Proof of concept

We conducted SDS-PAGE with (BBa_K2015012+BBa_K2015008/ BBa_K2015012+BBa_K2015009) on （pSB1C3）of E.coli（DH5α）. At first, we made the Gel for SDS-PAGE, with the following the Table1. What was the most important was to add TEMED ffinally because

Table. 1. Gel assay
Materials Volume
30％ Acrilmid 5 mL
Tris-Buffer 2.5 mL
SDS 100 µL
APS 100 µL
TEMED 5 µL
DW 2.295 ｍL
Total 10 ｍL


The following Table2 is about preparing the SDS-PAGE.

Table. 2. Experimental condition
Temperature IPTG Concentration (M) Volume (µL) Time to culture
37°C - - - 24 h
37°C + 0.4 6 24 h
37°C + 2 30 24 h
25°C - - - 16 h
25°C + 0.4 6 16 h
25°C + 2 30 16 h