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Table 2 Primer sequences and PCR reaction conditions for EVL/MIR342

From: Epigenetic silencing of miR-342-3p in B cell lymphoma and its impact on autophagy

  Forward primer (5′ to 3′) Reverse primer (5′ to 3′) Tm/cycles/MgCl2 Reference
(I) Methylation-specific PCR (MSP)
EVL/MIR342
  M-MSP AGG GAG CGT ATC GCG TTA C CGG AAA ACG CTA AAC TTA AAA CTA CG 55 °C/38x/2 mM NA
  U-MSP GGG AGG GAG TGT ATT GTG TTA T CTC CAA AAA CAC TAA ACT TAA AAC TAC A 57 °C/35x/1.5 mM NA
E-cadherin
  M-MSP GTG GGC GGG TCG TTA GTT TC CTC ACA AAT ACT TTA CAA TTC CGA CG 62 °C/40x/2 mM NA
  U-MSP GGT GGG TGG GTT GTT AGT TTT GT AAC TCA CAA ATA CTT TAC AAT TCC AAC A 58 °C/40x/2 mM NA
(II) Quantitative real-time reverse transcription-PCR
 EVL (NM_001330221.1) GCA GCG ACA ATG AGT GAA CA TGC TGA TCC TGC AAC TTG AC   
 DNMT1 GCT ACC TGG CTA AAG TCA AA CCA TTC CCA CTC TAC GG NA [30]
 E-cadherin GGA GGA GAG CGG TGG TCA AA TGT GCA GCT GGC TCA AGT CAA NA [31]
 GAPDH ACC ACA GTC CAT GCC ATC ACT TCC ACC ACC CTG TTG CTG TA NA [16]
(III) Cloning of luciferase reporter constructs with SRBS of miR-342-3p
 MAP1LC3B AAG GGC TAG CCC GCC TTT TTG GGT AGA AGT AGG GTC GAC AGT GAG GAC TTT GGG TGT GG 55 °C/35x/2 mM NA
  1. M-MSP methylated MSP, U-MSP unmethylated MSP, Tm annealing temperature, SRBS seed region binding site