Table 1 Primer sequences and PCR reaction conditions
From: Epigenetic silencing of a long non-coding RNA KIAA0495 in multiple myeloma
Primer set | Forward primer (5’ – 3’) | Reverse primer (5’ – 3’) | Product size (bp) | MgCl2/Tm/cycles | Reference |
|---|---|---|---|---|---|
(I) Methylation-specific polymerase chain reaction (MSP) | |||||
KIAA0495 | |||||
M-MSP | TGG AGA TAA CGG GTT TAG AGA AAT C | AAC GAA AAC AAA AAT AAA ACC TTC G | 191 | 1.5 mM/58 °C/40X | -- |
U-MSP | TGG AGA TAA TGG GTT TAG AGA AAT T | AAA CAA AAA CAA AAA TAA AAC CTT CA | 192 | 2.0 mM/58 °C/40X | -- |
(II) Reverse transcription-polymerase chain reaction (RT-PCR) | |||||
KIAA0495 | GCT GCT TGC TGT ACG TGG TG | CGT GGC TGA CAC AAA CTT GC | 178 | 1.5 mM/60 °C/35X | [16] |
GAPDH | ACC ACA GTC CAT GCC ATC ACT | TCC ACC ACC CTG TTG CTG TA | 452 | 1.5 mM/60 °C/24X | |