NlaIII NEB#R0125

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	Digest 164 bp products of PCR was completed at the condition of 5 units NlaIII (New England Biolabs) at 37°C for 4 hours. Restriction fragments were analyzed using 3% ethidium bromide-stained agarose gel.

Protocol tips
Digest 164 bp products of PCR was completed at the condition of 5 units NlaIII (New England Biolabs) at 37°C for 4 hours. Restriction fragments were analyzed using 3% ethidium bromide-stained agarose gel.

Publication protocol

Standard EDTA tubes were used to obtain blood samples, and DNA was obtained from each patient’s white blood cells using the QIAmp® DNA Blood Mini Kit. The blood samples were collected from patients during clinical evaluation. The DNA was extracted and analyzed by polymerase chain reaction (PCR) assay during the patient’s evaluation. After restriction fragment length polymorphism processing, IL-6 −174G/C polymorphism was analyzed using PCR (Figure 1). The primers for polymorphism analysis was used with the standard primers. Digest 164 bp products of PCR was completed at the condition of 5 units NlaIII (New England Biolabs) at 37°C for 4 hours. Restriction fragments were analyzed using 3% ethidium bromide-stained agarose gel. The 111- and 52-position fragments are consistent with the homozygous genotype C/C; 164 bp represents the homozygous genotype G/G. The three fragments in the positions of 111, 52, and 164 bp represent the heterozygous genotype G/C. The reliability of samples was analyzed and evaluated. A negative control was used to prevent false positive results.

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