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Table 1 Oligonucleotides used in this study

From: Analysis of RNA splicing defects in PITX2 mutants supports a gene dosage model of Axenfeld-Rieger syndrome

Name

Sequence*

Purpose

Ex-1b-f

GCGAATTCCAGTAGCCAAGGACTAGTAG

Forward and reverse primers to make exon 1b fragment

Int-1b-r

GCGGATCCAGAATTGCTCGCGCCCTTAG

 

Int-1b-f

GCGGATCCAGTGAATGTGCCGCTGCAGT

Forward and reverse primers to make exon 4 fragment

Int-4-r

GCGGTACCTCGGAGAGGGAACTGTAATC

 

Int-4-s

GCGGTACCTGGCTGAGTGATCAAACCGT

Forward and reverse primers to make exon 5 fragment

Ex-5-r

CGAAGCTTGGCGGCGCGTAAGGACAGG

 

IVS4+5G>C-f

GAGTCCGGGTAGcAGCCAGCACGGAG

Forward and reverse overlap PCR primers to make IVS4+5G>C mutant

IVS4+5G>C-r

CTCCGTGCTGGCTgCTACCCGGACTC

 

IVS5-11A>G-f

CTCCCTTGCCCCAgCCGCCCCCAGG

Forward and reverse overlap PCR primers to make IVS5-11A>G mutant

IVS5-11A>G-f

CCTGGGGGCGGcTGGGGCAAGGGAG

 

IVS4-1G>T-f

CGTTTTCAtAGAAAGAT

Forward and reverse overlap PCR primers to make IVS4-1G>T mutant

IVS4-1G>T-f

ATCTTTCTaTGAAAACG

 

T7 promoter

TAATACGACTCACTATAGGG

Forward primer to pcDNA T7 promoter to detect minigene mRNA

PCREx-1b-f

TGTCGGCCGTCTCCTCATCTTCC

Forward and reverse primers to detect endogenous and minigene mRNA

PCREx-5-r

TTGCGCTCCCTCTTTCTCCATTTG

 

GFP-f

GACGGCAACATCCTGGGGCACAAG

Forward and reverse primers to GFP

GFP-r

CGGCGGCGGTCACGAACTCC

 

GAPDH-f

TGATGACATCAAGAAGGTGGTGAAG

Forward and reverse primers to GAPDH

GAPDH-r

TCCTTGGAGGCCATGTGGGCCAT

 
  1. *Primers are 5' to 3', mutations are in lowercase