Device
106GFP

Part:BBa_K1679002

Designed by: Qikai Qin   Group: iGEM15_OUC-China   (2015-09-05)

GFP Reporter

This is a device containing a promoter (BBa_J23106),a RBS (BBa_B0034), a GFP coding sequence (BBa_E0040) and two terminators (BBa_B0010, BBa_B0012) on the pSB1C3, which can be easily transformed into E.coli and produce GFP (mut3b). This is a GFP expression device, which can express GFP (mut3b) without any induction.

Plate Reader Result

Equipment

Varioskan Flash Multimode Reader
Thermo Scientific
Read Speed: 6.4 well per second

Software

Run Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash
Current Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash

Protocol

Date: 2015 Aug 12
1. Set our instrument to read OD600
2. Setup a 96-well plate with our cultures
3. Take the measurement and record it
4. Calculate the dilution required for each sample (OD600=0.5)
5. Dilute each sample
6. Remeasure our sample on OD600
7. Recalculate our dilution and remeasure until it's within 5% of 0.5.

Unit

Three technical replicates of M9 liquid media was measured, which were called background value.
A series of concentration of sodium fluorescein was measured, which are 0, 10, 20, 40, 80, 160, 320, 640 ng/mL . A calibration curve was defined.

OUC-China-InterLab_2.jpg

Dataset

All samples were cut the mean of background value, and then compared with the calibration curve to get the final dataset in units of fluorescein.

OUC-China-InterLab3.jpg

OUC-China-InterLab_4.jpg

TR: Technical Replicate
BR: Biological Replicate

OUC-China-InterLab_5.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 705


[edit]
Categories
//classic/reporter
//function/reporter
//function/reporter/fluorescence
Parameters
chassisE. coli K-12 DH5-alpha
emission511nm
excitation501nm
function