Table 1 Designed primers for ARMS-PCR reaction
Description | General Characteristics |
|---|---|
ZAP70 | rs104893674 (Missense) |
Type of polymorphism | Single-base C > A |
Site of polymorphism | p.Ser518Arg |
PCR primers | |
Forward | 5- CGATGAAGGCCATGACCTC -3 |
Reverse A allele | 5- CTTCCGCAAGTTCTCCAGA -3 |
Reverse C allele | 5- CTTCCGCAAGTTCTCCAGC -3 |
Forward-PCR-Control | 5- CCTCTGCACAGTTTGGAC -3 |
Reverse- PCR-Control | 5- TCTGTCCAGCAATCCAGG -3 |
PCR conditions | |
Denaturation | 94 °C, 5 min |
Annealing | 58 °C, 40 s |
Extension | 72 °C, 40 s |
No. of cycles | 32 |
Product Size (bp) | 237 bp |
Beta-actin | Internal control |
PCR primers | |
Forward primer | 5- TATCCAGGCTGTGCTATCCCTGTAC -3 |
Reverse primer | 5- CTTGATGAGGTAGTCAGTCAGGTCC -3 |
PCR conditions | |
Denaturation | 94 °C, 5 min |
Annealing | 56 °C, 40 s |
Extension | 72 °C, 40 s |
No. of cycles | 32 |
Product size (bp) | 169 bp |