The Epoch BioTek is a monochronometer, which means it will measure any wavelength of absorbed or emitted light. You can choose up to six wavelengths to measure in a single assay. The software is Gen5 v 2.0. Use System Preferences to change where files are stored. We have free tech support. Protocols = sets of tasks including data reduction and data export. Experiments = uses protocols. The Take3 plate is a special plate that will read small quantities of samples (2 to 10 µl) or normal cuvettes, or special cuvettes designed to fit Take3 plate. Note, if using normal cuvettes, they need to have a cap, because they are inserted laying down. If you want to measure DNA using a regular 96 well plate, need to purchase special plates that are UV resistant
- Turn on computer and BioTek Epoch machine (it doesn’t matter which order).
- Open Gen5 software from computer
- Select "Read Now"
- Select Take3 application "Nucleic Acid Quantification"
- Select the following options
- Take3 Plate: Take3 (20161)
- Well Type: Microspots
- Sample Type: dsDNA
- Leave the Wavelengths box unchecked, unless you want more information
- Select “Blanks” from the drop-down menu on Well-type
- Select the microspots you will use for blanks by clicking on the circles
- If you are blanking all sixteen microspots proceed to Read
- Select "Samples" from the drop-down menu on well-type
- Select the microspots you will use for samples
- Wipe Take3 plate (microspots and glass) with a damp kim wipe to clean it
IMPORTANT: DO NOT DROP TAKE3 PLATE (it costs $1,000+). DO NOT REMOVE GLASS PLATE (it is factory measured for distance) DO NOT USE ALCOHOL (only clean plate with water or soap)
- The plate is held shut by a magnet, so hold plate down with finger while gently lowering lid closed.
- Place 3 µl of TE or water on the Take3 plate and 3 µl of your sample on the Take3 plate microspots. Alternatively you can just use TE on all the spots for an initial blanking. The spot will hold from 2 to 10 µl
- Gently close lid
- Select "Read", machine will make a little sound, door will open and window will tell you to put the plate in the tray
- Place the Take3 plate in tray and select "OK"
- Tray will close, machine will take the readings
- After reading, the program will show you the control CV’s and will state whether the CV’s are in an acceptable range or not. The CV measures the “Coefficient of variance” which basically tells you whether or not your multiple blanks are all giving similar readings. Generally you want to have a CV below 10% or > 0.10. The program refuses to give you any values until you click “accept” for the CV values. If the program finds really bad CV values it won’t let you continue
- If you have run both blanks and samples on the same plate, the program will open excel and show you the readings for your samples. You can save or print this file, or just write down the numbers. If you have more samples, return to the Gen5 program and read the next plate.
- The program will continue to add excel worksheets to the spreadsheet file until you click End of Batch.
- When you click End of Batch the program will create a summary spreadsheet on the excel spreadsheet.
- Save the excel spreadsheet or print it.