HLA SSP 96 Well Primer Position
Primer pairs are designed for typing the HLA class I genes A (24 primer pairs), B (48 primer pairs), C (16 primer pairs) and the class II genes DQB (8 primer pairs), DRB1 (21 primer pairs), DRB3(1 primer pair), DRB4(1 primer pair), DRB5(1 primer pair). For the combined testing of ABDRDQ the number of DRB1 and DQB primer pairs was reduced to 19 and 5 respectively. The PCR accomplished with these primers produce well-defined DNA fragments of different length. The Primer-Mixes also contain in addition to the Sequence Specific Primers (SSP) control primers of the human globin gene at a lower concentration. They act as an internal control for the PCR. They show if the PCR reaction had taken place and to exclude results based on inhibitors, non-sufficient DNA and displaced primers from other wells. For the PCR of the class I genes the length of the fragments is 1070 bp, and for PCR of the class II it is 429 bp. The amplified DNA fragments will be detected in an agarose gel by dyeing the double stranded DNA with ethidium bromide.
