Primers were designed via Methy Primer Express v1.0 (Applied Biosystems, USA) and synthesized in Sangon Biotech (Shanghai, China).The primers are shown in Table s1 . In total, 1.5 µl bisulfite-treated DNA were amplified in a 30 µl reaction mixture consisting of 1 × PCR Buffer with 0.5U AmpliTaq Gold DNA polymerase (Applied Biosystems, USA), 0.2 mM dNTP mix, and 0.3 µM of each primer. The PCR conditions were as follows: 95 °C for 10 min, then 40 cycles of 95 °C for1 min, 55 °C for 1 min, 72 °C for 1 min and finally an elongation step of 7 min at 72 °C. The PCR products were separated on a 2% agarose gel, prestained with Gelred (Shanghai Life iLab Bio, China) and visualized by UV transillumination. Leukocyte (leu) DNA from healthy women was used as a negative control, and in vitro methylated (iv) leukocyte DNA was used as a positive control.
Methy primer express v1
Manufactured by Thermo Fisher Scientific
Sourced in United States
Methy Primer Express v1.0 is a software application designed for the design of methylation-specific PCR (MSP) primers. The core function of the software is to help researchers identify appropriate primer sequences for the detection of DNA methylation patterns.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using methy primer express v1
1
Bisulfite-treated DNA Amplification Protocol
2
Methylation-Specific PCR Protocol
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Primers were designed via Methy Primer Express v1.0 (Applied Biosystems, USA) and synthesized in Sangon Biotech (Shanghai,China).The primers were shown in Table S2. In total, 1.5ul bisul te-treated DNA were ampli ed in a 30 ul reaction mixture consisting of 1x PCR Buffer with 0.5U AmpliTaq Gold DNA polymerase (Applied Biosystems,USA), 0.2 mM dNTP mix, and 0.3 uM of each primer. The PCR conditions were as follows: 95 °C for 10 min, then 40 cycles of 95°C for 60 s, 55°C for 60 s, 72°C for 1 min and nally an elongation step of 7min at 72°C. PCR products were separated on a 2% agarose gel, prestained with Gelred(Shanghai Life iLab Bio,China) and visualized by UV transillumination. Leukocyte (leu) DNA from healthy women was used as a negative control, and in vitro methylated (iv) leukocyte DNA was used as a positive control.
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