Figure 4

Diagram of the typing procedure. In 4A, in the first phase, allele specific primers are shown in red and black. The sample is shown as wavy lines in grey and blue. There is a PCR reaction in the supernatant with forward and reverse allele-specific primers. The PCR amplicon is then kidnapped and attaches to one of the reverse primers on the solid phase. In the presence of nucleotides and polymerase a second chain is formed. In 4B, the kidnapped amplicon, which is only held by hydrogen bonding, is released by heating. Washing leaves a covalently bonded single strand. In 4C, a reverse primer is introduced which binds to the covalently attached amplicon. In reality we have all possible forward and reverse primers illustrated as 3 candidate SNP in a row in green, red and grey. In 4D, the results from the separate reactions can be combined to indicate the SNP haplotypes.