---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2010-09-18 03:57:46+00:00
layout: post
slug: opticon-calibration
title: Opticon Calibration
categories:
  - 2010
  - Miscellaneous
tags:
  - calibration
  - FAM
  - graphs
  - Opticon2
  - troubleshooting
---

Distributed 50uL of FAM calibration dye to wells. Ran out of dye!!

Looking back at old purchasing logs, it turns out we need 2 orders of dye packs to have enough for a 96-well plate.

Will cap existing plate with dye, wrap in foil and store @ -20C.

Ordered an additional pack of dye (Cat# 10006046; not available online, must call BioRad to order). Will ship on Monday. Will finish calibration procedure on Tuesday (20100928). Ugh.

Results:

Empty Plate:

![](https://eagle.fish.washington.edu/Arabidopsis/20100917%20Opticon%20Cal%20Empty.JPG)

Plate with dye (presumably calibrated):

![](https://eagle.fish.washington.edu/Arabidopsis/20100922%20Opticon%20Cal%20Dye.JPG)

After running the calibration protocol with the dye, all the wells should show consistent fluorescence levels. Clearly, they do not. Oddly enough, there appears to be a cyclical pattern across the wells of low -> high -> low fluorescence. The calibration protocol advises that if the wells do not exhibit consistent fluorescence across wells, then the plate should be read again. The graph above is the 2nd reading, which appears to be the same as the 1st. Conclusion is that the Opticon 2 is not working correctly and will contact BioRad for price quotes on repairs.