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Table 5 Oligonucleotide primer combinations and PCR conditions (previously not described) used for PCR-ASP in this study.

From: Two previously proposed P1/P2-differentiating and nine novel polymorphisms at the A4GALT (Pk) locus do not correlate with the presence of the P1 blood group antigen

Primer combination

Specific primer (pmol)

Control primer

DMSO (%)

Annealing temperature (°C)

Extension time (sec)

  

HGH-F/R (pmol)

JK-L-F/R (pmol)

MO-21/31 (pmol)

   

Pk-5'-(-1056)-F

5

1

   

64

40

Pk-5'-(-550)-R

5

      

Pk-5'-(-1056)F

5

1

   

64

40

Pk-5'-(-550insC)-R

5

      

Pk-5'-(-834)-F

7.5

 

0.5

 

4

62

60

Pk-5'-(-160A-mis)-R

7.5

      

Pk-5'-(-834)-F

5

 

0.5

 

3

62

60

Pk-5'-(-160G-mis)-R

5

      

Pk-5'(-550insC)-F

10

  

0.75

 

64

40

Pk-5'-(-160G)-R

10

  

0.75

   

Pk-5'(-550T)-F

7.5

  

0.5

 

63

40

Pk-5'-(-160A)-R

7.5

  

0.5

   

Pk-109A-F or Pk-109G-F

6

0.4

  

2

65

90

Pk-1697A-R or Pk-1697G-R*

6

      

Pk-987A-F

5

0.4

   

66

60

Pk-1697A-R

5

      

Pk-987G-F

5

0.4

   

66

60

Pk-1697G-R

5

      

Pk-109A-F or Pk-109G-F

7.5

 

0.5

  

64

60

Pk-1120-R**

       
  1. Denaturation was carried out at 96°C for 7 min followed by 35 cycles at 94°C for 30 s. The annealing time was 30 s.
  2. * Four double allele-specific amplifications were performed.
  3. ** These combinations were used for allele-specific amplifications for sequencing purposes.