Kidd Lab 

NlaIII site (functional polymorphism in exon 4)

Template: Genomic DNA 300 ng/reaction volume of 25 µl

Primer 1.    Forward Primer:    5'-GCC CGC CTG CTG TCA CC-3'                  Final Concentration = 200 nM
Primer 2.    Reverse Primer:    5'-CTG AGG GGC CTG GTG ATA GTG-3'        Final Concentration = 200 nM

dNTPs: Final Concentration = 200 µM each

Buffer: Final Concentration 1.5 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4

Taq DNA Polymerase: 1.0 Units/reaction (Perkin Elmer AmpliTaq DNA polymerase)

PCR Cycling Profile: 94^oC (1')
                              94^oC (30"), 60^oC (30"), 72^oC (30")  × 30 cycles
                              72^oC (10')

Product Size:
                               NlaIII
                   238 bp -------------> 114 bp COMT*H, uncut allele, or 96 bp and 18 bp, COMT*L allele, cut allele.
                                              Constant bands: 70 bp and 54 bp
                                              (site absent is Val; site present is Met)

Reference:
Chen CH, Lee YR, Wei FC, Koong FJ, Hwu HG, Hsiao KJ. 1997.  Association study of NlaIII and MspI genetic polymorphisms of catechol-O-methyltransferase gene and susceptibility to schizophrenia.  Biol Psychiatry 41(9): 985-987
 
 

Kidd Lab Histograms for Allele Frequencies