Comparison of Staining Dyes

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Revision as of 23:55, 27 April 2010 by >Jbrubake (Created page with '=Purpose= To establish the best dye to use for any subsequent experiments in order to maintain consistency in results. =Methods= 3 dyes were tested: Dapi, SYBR Green, and YOYO…')
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Purpose

To establish the best dye to use for any subsequent experiments in order to maintain consistency in results.

Methods

3 dyes were tested: Dapi, SYBR Green, and YOYO. Each dye was diluted to 2X concentration.

Sample tubes 1 and 2 were combined, filtered through a 5um filter, and used as a standardized sample.

For each dye:

- Use a clean 0.2mL tube
- Add 7uL sample + 3uL 2X dye
- Incubate for 10 minutes in the dark
- Add 10uL water
- Vortex and spin down

Each sample was plated and imaged under a fluorescence microscope using 10X magnification.

Results

File:Dapi 10X Comparison.jpg

Image 1: 2X Dapi

File:SYBR Green 10X Comparison.jpg

Image 2: 2X SYBR Green

[[image: YOYO_10X_Comparison.jpg

Image 3: 2X YOYO