The SNPs were genotyped by using primer extension of multiplex products with detection by matrix-assisted laser desorption ionization – time of flight mass spectroscopy using a MassARRAY Compact Analyzer (Sequenom, San Diego, CA, USA).
Multiplex products (1 : 140 dilutions) were genotyped using an ABI 3730 sequencer.
For the fragment analyses, 1 μL of 1 20 diluted multiplex products was used.
All genotyping was done using the primer extension of multiplex products with detecting by matrix-assisted laser desorption ionization time of flight mass spectroscopy using a MassARRAYCompact Analyzer (Sequenom, San Diego, CA, USA).
All genotyping was done using the primer extension of multiplex products with detecting by matrix-assisted laser desorption ionization-time of flight mass spectroscopy using a MassARRAY Compact Analyzer (Sequenom, San Diego, CA, USA).
All the SNPs were genotyped using the primer extension of multiplex products with detection by matrix-assisted laser desorption ionization-time of flight mass spectroscopy using a MassARRAY Compact Analyzer (Sequenom, San Diego, CA, USA) and the overall call rate was 98.7%.
For de novo replication in stage 2, all selected SNPs were genotyped in the Hong Kong and Shanghai case control samples by a primer extension of multiplex products with detection by Matrix-assisted laser desorption ionisation-time of flight mass spectroscopy using a MassARRAY platform (Sequenom; San Diego, CA, USA).
Apparatus and protocol for characterizing online DNA and YOYO-1 interesolution refficiency efficiency, and retention time varying with number of runs; preparation of chip injector; capillary surface modification for capillary PCR; capillary PCR; incompatibility of YOYO-1 with PCR; and calculation of the three multiplex products length are provided as Supporting Information.
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In a second round of PCR, each TRPC subunit and hcrt/orx cDNAs were specifically amplified (using 1 µM of each TRPC nested primers and 0.1 µM of each hcrt/orx primers) in individual reactions with 2 µl of the multiplex PCR product under similar conditions for 35 cycles (94°C for 30 s, 52°C for 45 s, 72°C for 2 min).
