ZEG Genotyping Best Practices Guide

ZEG User Manual

Watch the ZEG User Video!

Download the ZEG White Paper and learn how to accelerate your research discoveries!

ZEG Development

The Zebrafish Embryo Genotyper was conceptualized, validated, published and patented at the University of Utah by the Department of Mechanical Engineering and Pediatrics. Graduate students who lead this development were Christopher Lamber, Briana Fresher, Arien Chung, et al. Read and cite the research article in your Zebrafish ZEG experiments.

CITATION GUIDE

Frequently Asked Questions

How do I increase the quality of the extracted DNA using PCR?

The polymerase/mix you use is critical. Best results are obtained using Phire Polymerase, #F126 from Thermo-Fisher, an amplicon smaller than 300bp and 40 cycles of PCR.

How do I increase the quality of the extracted DNA post-PCR using HRMA?

For HRMA, the best results use LC Green from Bio Fire (you need to optimize the annealing temp; it is usually a bit higher) or Bio Rad’s HRMA master mix.

How do I increase the concentration of the extracted DNA using PCR?

To increase the concentration extracted we suggest not to dilute or do a Thermopol isolation on the ZEG samples at all, use 5 ul of ZEG material directly in the PCR mix.

Are chips Reusable?

It is not recommended to reuse chips; this will lead to cross contamination if not cleaned properly.

Do I have to optimize my PCR before using the ZEG?

We recommend optimizing your PCR procedure before you use the ZEG. Design primers that amplify more robustly and reliably on a fin clip, and then try them on ZEG.

Can I use the ZEG on unhatched Zebrafish Embryos?

Fish inside the chorion won’t work. If you must do ZEG before they hatch, manually dechorionated them first, or wait until they hatch.

Do I need to use mineral oil for PCR or HRMA?

You don’t need mineral oil for PCR but you do need it for HRMA.

How many PCR samples can I get from one run?

If you use 15ul for each embryo on the chip, you get back at least ~12ul. You could do potentially 3 PCRs with that, using 4ul each.

What is the voltage on the ZEG supposed to be set too?

All ZEG units manufactured and sent since July 2022 are outside the range ZP-001 and ZP-041, so the voltage should be set to 1.8V. Those sent earlier to 1.4 V.

How do I change the ZEG voltage?

If you need to change the voltage:

The base unit controller voltage can be changed from 1.4 V to 2.4 V by the following method. PLEASE NOTE: Do not set the voltage to or beyond 3V. This will damage the vibration motor.
Press the yellow SET button. This would highlight the ‘SET’ word on the LED screen.
Press in the silver knob several times until the ‘1’ of the ’01.80’ (on the LED screen) is highlighted.
Rotate the knob clockwise (to increase) or counter-clockwise (to decrease) this value. Change this value from ‘1’ to ‘2’, until desired voltage is set.
Press in the silver knob.
Press the yellow SET button until the screen returns to original screen.

Is it possible to add anesthesia to the extraction water without having any impact on the results?

Tricaine (the typical fish anesthesia) doesn’t affect ZEG results in any way.

Is the cover enough to avoid evaporation or are there any extra precautions we should follow?

There aren’t any extra precautions you need to take to avoid evaporation, the cover provided with the ZEG is enough.

I have faulty chips, what do I do?

Please contact customer support by emailing info@wfluidx.com

My ZEG stopped working, what do I do?