Difference between revisions of "Part:BBa K1679001"

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Revision as of 00:59, 19 September 2015

GFP Reporter

This is a device containing a promoter (BBa_J23101),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.

Layout

101: J23101+I13504; 106: J23106+I13504; 117: J23117+I13504; 117R: J23101+Ribo J+I13504; D: E. coli K-12 DH5α without plasmid; R: R0040; I: I0270; 1: Technical replicate 1; 2: Technical replicate 2; 3: Technical replicate 3; A:Biological replicate 1; B: Biological replicate 2; C: Biological replicate 3; M9: M9 liquid media; \: Blank control'SF: Sodium Fluorescein (concentration of 0, 10, 20, 40, 80, 160, 320, 640 ng/mL)

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.

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.

TR: Technical Replicate
BR: Biological Replicate

Sequence and Features

Functional Parameters