Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Aug 13;10:13757. doi: 10.1038/s41598-020-70735-2

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Identification of heterozygous F1 animals with CRISPR/Cas9 indel mutations of the mtnr1a gene. (a) Representative RFLP analysis shows that PCR products of mtnr1a VIL heterozygotes display two predicted cleavage bands (arrows), whereas WT and non-VIL mutant DNA is not cleaved. The creation of a BseRI site by the VIL deletion is illustrated in Supplementary Fig. S3. (b) Direct PCR Sanger sequencing chromatogram of F1 WT progeny display prominent peaks for all bases in the T2 sgRNA target area. The T2 PAM sequence (CCT) is boxed in black, and the downstream 9-bp sequence of the mtnr1a VIL deletion mutation is boxed in blue. The corresponding amino acid sequence is shown below the nucleotide codon sequence. (c) In an F1 tadpole heterozygous for the mtnr1a 9-bp (VIL) deletion, the trace decomposes (i.e., mixed base calls) at the predicted target site 4-bp downstream of the T2 PAM site, and continues to the end of the trace, as expected for a heterozygous indel mutation. The decomposed sequence represents the overlapping peaks of the mtnr1a WT and VIL alleles. (d) Partial view of alignments from Poly Peak Parser (https://yosttools.genetics.utah.edu/PolyPeakParser/) output revealing the heterozygous 9-bp mtnr1a VIL deletion mutation. The WT allele is the top line and the mutant allele is the bottom line.