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Oct 16, 2019

Version 2

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# Microplate Reader Workflow V.2

DOI

https://dx.doi.org/10.17504/protocols.io.8bqhsmw

NUS iGEM¹

¹National University of Singapore

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NUS iGEM

National University of Singapore

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DOI: https://dx.doi.org/10.17504/protocols.io.8bqhsmw

**Protocol Citation:** NUS iGEM 2019. Microplate Reader Workflow. protocols.io

https://dx.doi.org/10.17504/protocols.io.8bqhsmw

**License:** This is an open access protocol distributed under the terms of the **Creative Commons Attribution License**, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

**Protocol status:** Working

We use this protocol and it's working

Created: October 16, 2019
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Last Modified: September 12, 2023

Protocol Integer ID: 28752

Troubleshooting
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1. Refresh overnight cell culture in LB media at 37 °C

Protocol

NAME

Preparation of LB Media

CREATED BY

NUS iGEM

Preview

1.1. Weigh **燃燒 25 g** of Luria Broth Base powder.

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Luria Broth Base (Miller's LB Broth Base)&trade;, powder Thermo
Fisher Catalog #12795027

1.2. Add the powder into **1 L** of water.

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Water refers to sterilized deionized water

1.3. Autoclave entire bottle of LB media.

2. Measure OD₆₀₀ of cell culture for desired OD value

3. Load **200 µL** of cell samples into three wells of a 96-well plate (triplicates)

4. Induce appropriate volumes of chemical inducers (not exceeding **6 µL**) in each well

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5. Include LB media as blanks
6. Load the plate (with lid) into microplate reader

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7. Start protocol to run continuous measurement overnight
Note

**OD<sub>600</sub> (absorbance) protocol:**

Wavelength: 600nm

Measurement interval time: 8min

Run time: 18h

**GFP (fluorescence) protocol:**

Excitation: 485 +/- 10

Emission: 528 +/- 10

Gain: Extended

**RFP (fluorescence) protocol:**

Excitation: 535 +/- 20

Emission: 600 +/- 20

Gain: Extended

**Luminescence protocol:**

Gain: 118

Integration time: 1s

**OD<sub>600</sub> + GFP protocol script:**

Measurement interval time: 8min

Run time: 18h

**OD<sub>600</sub> + luminescence protocol script:**

Measurement interval time: 20min

Run time: 20h