Data from: Reliable and fast genotyping protocol for galactosylceramidase (GALC) in the Twitcher (Twi) mouse

The Twitcher (Twi) mouse is a neurological Krabbe disease (KD, or globoid
cell leukodystrophy) spontaneous mutant line. The genome of the Twi mouse
presents a single nucleotide polymorphism (SNP), leading to an
enzymatically inactive galactosylceramidase (GALC) protein and causing KD.
In this context, mouse Twi genotyping is an essential step in KD research.
To date, the genotyping method used is labor-intensive and often
drives ambiguous results. Here, we evaluate a novel protocol for
the genotype determination of GALC mutation status in Twi mice based on
the allele-discrimination real-time polymerase chain reaction (PCR).DNA is
extracted from Twi mice (n = 20, pilot study; n = 120, verification study)
and control group (n = 10, pilot study; n = 30 verification study) and
assessed by allele-discrimination real-time PCR to detect SNP
c.355G>A.Using the allele-discrimination PCR, all the samples are
identified correctly with the genotype GG (wild-type, WT), GA
(heterozygote, HET), or AA (homozygote, HOM) by the first analysis and no
animals were not genotyped.We demonstrated that this novel method can be
used to distinguish timely, accurately and without ambiguity
among HOM, WT, and HET animals. This protocol represents a great
opportunity to increase accuracy and speed in KD research.