Delayed Diagnosis, Difficult Decisions: Novel Gene Deletion Causing X-Linked Hypophosphatemia in a Middle-Aged Man with Achondroplastic Features and Tertiary Hyperparathyroidism

Table 2

Genomic DNA isolation and mutation screening, DNA extraction, and amplification of PHEX exons by PCR.

PCR thermocycling conditions for amplifying the PHEX gene		Forward and reverse primer sequences for screening exon 14 [4]
Steps	Conditions
Initial denaturation	95°C for 3 minutes	Forward primer	CAACA AGTAGGTGAC TGTCGAGCC (NG_007563.2, 144846–144869)
35 cycles		Reverse primer	GCTAAG CTATGAGGAC ACAAAGGC (NG_007563.2, 147203–147226)
Denaturation	95°C for 30 seconds
Annealing	60°C for 45 seconds
Extension	72°C for 60 seconds
Final extension	72°C for 10 minutes
Hold	4°C

Annealing temperature is specific to each primer set; 60°C for screening exon 14 of PHEX.