Team:SDU CHINA/interlab

Background

“All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology.” Our team took part in this study which aimed to be one of the labs to repeat measurements in different condition. In our case, we measured fluorescence using plate reader.

Design

We often used fluorescence as an indicator of promoter activity. However, the ability to repeat measurements in different labs has been difficult. To solve this problem, InterLab study has been developed as a robust measurement procedure for green fluorescent protein (GFP) which is one of the most used markers in synthetic biology.

This year's iGEM Interlab study is to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable and compare the results from various teams. iGEM Interlab study has provided 2 protocols of measuring GFP, and we choose “plate reader”. We have to measure absorbance at 600 nm and GFP fluorescence for GFP expression per cell for all devices.

Materials

Plasmids used

• Positive Control Device: I20270 in pSB1C3 (Located in Kit Plate 6, well 20B)
• Negative Control Device: R0040 in pSB1C3 (Located in Kit Plate 6, well 20D)
• Test Device 1: J23101+I13504 in pSB1C3 (Located in Kit Plate 6, well 20F)
• Test Device 2: J23106+I13504 in pSB1C3 (Located in Kit Plate 6, well 20H)
• Test Device 3: J23117+I13504 in pSB1C3 (Located in Kit Plate 6, well 20J)
• Test Device 4: J23101.BCD2.E0040.B0015 in pSB1C3 (Located in Kit Plate 6, well 20L)
• Test Device 5: J23106.BCD2.E0040.B0015 in pSB1C3 (Located in Kit Plate 6, well 20N)
• Test Device 6: J23117.BCD2.E0040.B0015 in pSB1C3 (Located in Kit Plate 6, well 20P)