Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Xing, Lingyan; Quist, Tyler S.; Stevenson, Tamara J.; Dahlem, Timothy J.; Bonkowsky, Joshua L.

doi:10.3791/51138

Back to Previous Page

- Add this to...
- Twitter
- Facebook
- Google Bookmarks
- LinkedIn

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Published Date:

Feb 05 2014

Publisher's site:

http://dx.doi.org/10.3791/51138 New Site Icon

Source:

J Vis Exp. 2013; (84):e51138.

[PDF-607.89 KB]

Details:

Personal Authors:

Xing, Lingyan ; Quist, Tyler S. ; Stevenson, Tamara J. ; Dahlem, Timothy J. ; Bonkowsky, Joshua L. ;

Keywords:

[+]

Description:

Zebrafish is a powerful vertebrate model system for studying development, modeling disease, and performing drug screening. Recently a variety of genetic tools have been introduced, including multiple strategies for inducing mutations and generating transgenic lines. However, large-scale screening is limited by traditional genotyping methods, which are time-consuming and labor-intensive. Here we describe a technique to analyze zebrafish genotypes by PCR combined with high-resolution melting analysis (HRMA). This approach is rapid, sensitive, and inexpensive, with lower risk of contamination artifacts. Genotyping by PCR with HRMA can be used for embryos or adult fish, including in high-throughput screening protocols.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

DP2 MH100008/DP/NCCDPHP CDC HHS/United States
DP2 MH100008/MH/NIMH NIH HHS/United States
R01 MH092256/MH/NIMH NIH HHS/United States

Supporting Files:

	nihms671211.nxml	text/plain
	nihms671211f1.gif	image/gif
	nihms671211f1.jpg	image/jpeg
	nihms671211f2.gif	image/gif
	nihms671211f2.jpg	image/jpeg
	nihms671211f3.gif	image/gif
	nihms671211f3.jpg	image/jpeg

No Related Documents.

You May Also Like:

Microfluidic-aided genotyping of zebrafish in the first 48 h with 100 % viability

Personal Author:

Samuel, Raheel; Stephenson, Regan; Roy, Paula; Pryor, Rob; Zhou, Luming; Bonkowsky, Joshua L.; Gale, Bruce K.;

Published:

Apr 2015

Source:

Biomed Microdevices. 17(2):43.

Description:

This paper introduces an innovative method for genotyping 1-2 days old zebrafish embryos, without sacrificing the life/health of the embryos. The method utilizes microfluidic technology to extract and collect a small amount of genetic material from t...

File Type:

[PDF - 1.07 MB]
Nitroreductase-Activatable Morpholino Oligonucleotides for in Vivo Gene Silencing

Personal Author:

Yamazoe, Sayumi; McQuade, Lindsey E.; Chen, James K.;

Published:

Jul 28 2014

Source:

ACS Chem Biol. 2014; 9(9):1985-1990.

Description:

Phosphorodiamidate morpholino oligonucleotides are widely used to interrogate gene function in whole organisms, and light-activatable derivatives can reveal spatial and temporal differences in gene activity. We describe here a new class of caged morp...

File Type:

[PDF - 1.40 MB]
Real-time PCR assays for genotyping of Cryptococcus gattii in North America

Personal Author:

Kelley, Erin J; Driebe, Elizabeth M; Etienne, Kizee; Brandt, Mary E; Schupp, James M; Gillece, John D; Trujillo, Jesse S; Lockhart, Shawn R; Deak, Eszter; Keim, Paul S; Engelthaler, David M;

Published:

May 13 2014

Source:

BMC Microbiol. 2014; 14:125.

Description:

Background : Cryptococcus gattii has been the cause of an ongoing outbreak starting in 1999 on Vancouver Island, British Columbia and spreading to mainland Canada and the US Pacific Northwest. In the course of the outbreak, C. gattii has been identif...

File Type:

[PDF - 649.39 KB]
A General Method for Regulating Protein Stability with Light

Personal Author:

Bonger, Kimberly M.; Rakhit, Rishi; Payumo, Alexander Y.; Chen, James K.; Wandless, Thomas J.;

Published:

Nov 08 2013

Source:

ACS Chem Biol. 9(1):111-115.

Description:

Post-translational regulation of protein abundance in cells is a powerful tool for studying protein function. Here, we describe a novel genetically encoded protein domain that is degraded upon exposure to nontoxic blue light. We demonstrate that fusi...

File Type:

[PDF - 403.86 KB]
The Evolution of Advanced Molecular Diagnostics for the Detection and Characterization of Mycoplasma pneumoniae

Personal Author:

Diaz, Maureen H.; Winchell, Jonas M.;

Published:

Mar 08 2016

Source:

Front Microbiol. 2016; 7.

Description:

Over the past decade there have been significant advancements in the methods used for detecting and characterizing Mycoplasma pneumoniae, a common cause of respiratory illness and community-acquired pneumonia worldwide. The repertoire of available mo...

File Type:

[PDF - 904.79 KB]