Coding Region Mutation Screening in Optineurin in Chinese Normal-Tension Glaucoma Patients

Table 1

Oligonucleotide primers used for PCR and sequencing of OPTN in NTG patients.

	Primer	Sequence (5 to 3)	Amplicon Size (bp)	MgCl2 (mM)	Anneal Temp (°C)
1	OPTN-3/4F	TTTCTGAAGCTACATATACCTTT	445	2.5	62-55 (TD)
	OPTN-3/4R	CTCACCACCAAGCCTTAGTG
2	OPTN-5F	AGCCATGTGGTCAAGTGGAC	362	1.5	62-55 (TD)
	OPTN-5R	AAGATTCCACTGCAAATGTCAA
3	OPTN-6F	GTGCCCAGCCTTAGTTTGATC	336	1.5	58
	OPTN-6R	AATCCTTGGCTTGTGTTGACA
4	OPTN-7F	TTGTGTTAAATCCCTTGCATTT	202	1.5	55
	OPTN-7R	CCTCAGGTCACAACATTTGAC
5	OPTN-8F	CAGTTCCTTTAAGCTGGTCC	381	1.5	58
	OPTN-8R	TTGCACAAGCCTGGAATATTC
6	OPTN-9F	CCTGATCCTTTATCCCAATTGT	244	1.5	58
	OPTN-9R	GTAGTGGCCGTGGAATCTTA
7	OPTN-10F	GGCTACTAATGGTTCAGCCTG	239	1.5	62-55 (TD)
	OPTN-10R	GGAGGATAAAATTGCTCTCTCA
8	OPTN-11F	TGCGACGTAAAGGAGCATTG	230	1.5	62-55 (TD)
	OPTN-11R	TCGCTGCCCTTCTGACTCAA
9	OPTN-12F	TGGGAGGCAAGACTATAAGTTT	221	1.5	55
	OPTN-12R	CGTTCAACAGTTTCTGTTCATT
10	OPTN-13F	CAGGCAGAATTATTTCAAAAC	273	2.5	62-55 (TD)
	OPTN-13R	TTTCCAATGCGAGAATACAG
11	OPTN-14F	CTCCTCATCGCATAAACACTGT	267	1.5	58
	OPTN-14R	GGCACCTCTTCTACGGCC
12	OPTN-15F	ACTTCTGTGGACTGTCTGCTC	263	1.5	62-55 (TD)
	OPTN-15R	TGATTTGGAATCCATTGTAGAG
13	OPTN-16F	TCGCCATCTGTTCTTCAAGT	272	1.5	58
	OPTN-16R	CACAAAAGCACAACTCTTGGA

TD: touchdown PCR using a cycling program where the annealing temperature is gradually reduced by 0.2°C per cycle.