Team:BNDS China/Results

Results

Here are the transformation of Orai1 V102A and pGEX-kg:

fig.1 The bacteria(Orai 1 V102A transformed in E. coli TOP10) growth on the LB media + ampicillin Oct. 4, 2016

fig.3 The bacteria(Orai 1 V102A transformed in E. coli TOP10) growth on the LB media + ampicillin Oct. 5, 2016

fig.2 The bacteria(Orai 1 V102A transformed in E. coli TOP10) growth on the LB media + ampicillin Sep. 16, 2016

fig.4 The bacteria(Orai 1 V102A transformed in E. coli TOP10) growth on the LB media + ampicillin Sep. 16, 2016

Here is the result of plasmid extraction of pGEX-kg and the result of Orai1 V102A plasmid undergoes Polymerase Chain Reaction.

fig.5 The plasmid pGEX was extracted from the bacteria cultured and the PCR was successful.

By comparing the size of the round plasmid and the linear DNA that has cut by enzymes, we inferred that our digestion has succeed.

The result of transformation after ligation:

fig.7 Bacteria after liagtion of pSB1C3 and Orai1 and transformation have grown on the LB plate adding chloromycetin. We labeled them on the back of the plate and used colony PCR to test the result. (fig.9) The “massive” result makes us nervous.

fig.8 Bacteria after liagtion of pGEX and Orai1 V102A and transformation have grown on the LB plate adding ampicillin. We labeled them on the back of the plate and used colony PCR to test the result. (fig.10)

The result of colony PCR Test:

fig.9 The electrophoresis testing result of colony PCR (pSB1C3-Orai1). The marker and the bands suggest that we had successfully connect two of them!!!

fig.10 The electrophoresis testing result of colony PCR (pGEX-Orai1). The last six colony numbers all have bands at about 700 (the length of our part). We still need to do sequencing to confirm.

The result of colony PCR Test:

fig.11 The bacteria(pSB1C3-Orai1) growth on the LB media + chloromycetin Oct. 16, 2016

fig.12 The bacteria(pGEX-Orai1) growth on the LB media + ampicillin Oct. 16, 2016

fig.13 the result after the final plasmid undergoes PCR. According to the graph, the first one has successfully connected.