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Table 1 Quantitative RT-PCR primer sequences and reaction conditions for CDKN1C.

From: Evolution of the CDKN1C-KCNQ1 imprinted domain

Primers

 

Fw primer (5'to 3')

GCCTCAAACCCTTTCACCT

Rv primer (5'to 3')

CGCTTACGGGTCCTCTGAT

Reagents

 

MasterMix

10 ul, 2×

Primer mix

150 uM final each

H2O

4.1 ul

cDNA

5 ul, ~5 ng

Conditions

 
 

50°C 10 min

 

95°C 15 min

39 ×

95°C 30 sec

 

60°C 20 sec

 

72°C 40 sec

 

78°C 1 sec

 

Plate read

 

72°C 1 min

Melting curve

55 – 90°C, 0.5°C, 1 sec

 

72°C 5 min

 

15°C Hold

  1. Melting curve analyses were performed after each PCR and one sample from each triplicate was assessed by gel electrophoresis to confirm there was no contamination.