Journal: Experimental Animals
Article Title: Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis
Figure Lengend Snippet: Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.
Article Snippet: The difference between homozygous (Hrx /Hrx ) and wild-type (Hr /Hr ) genomes was determined using multiple PCRs with HotStarTaq (#203443, Qiagen; regions 1, 8–13) or KOD FX neo (KFX-201, TOYOBO, Osaka, Japan; regions 2–7) DNA polymerases.
Techniques: Mouse Assay, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis