Appendix 2 of Manes, Mol Vis 2017; 23:198-209.


Appendix 2. Sequence of the primers (5'-3'), fusion temperatures and concentration of MgCl2 used to amplify GUCA1A (NM_000409.3)

To access the data, click or select the words “Appendix 2.” Amplification condition was performed in a 25 µl reaction volume and consisting of 16.8 µl nuclease free water, 2.5 µl of TAQ polymerase buffer, 1.5 or 2 µl MgCl2, 0,5 µl forward primer, 0,5 µl reverse primer, 0.2 µl TAQ polymerase, 2 µl template. Thermal cycle programmed for 90 s at 95 °C as initial denaturation, followed by 35 cycles of 30 s at 95 °C for denaturation, 30 s at 60 °C or 62 °C as annealing, 60 s at 72 °C for extension, and final extension at 72 °C for 10 min. PCR products were examined by electrophoresis at 100 V for 30 min in a 1.5% (w/v) agarose gel in 1X TAE buffer. The marker used DNA ladder 100 bp. Electrophoresis gel was soaked in ethidium bromide for 30 min then visualized in UV light.