Skip to main content

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