GPC1, HAS1, HAS2, HAS3, EXT1, EXT2, DEFB1, TJP1, and CLDN3 expression was quantified with SYBR® Green Real time PCR Master Mix (Sigma). Reactions were carried out as previously described (5 (link)). The 2−ΔCt method was used to calculate relative mRNA expression normalized to the housekeeping gene GAPDH. The 2−ΔΔCt method was used for calculating fold change in gene expression levels versus untreated controls. All the primers were synthesized by Sigma-Aldrich as previously described in
Protocol detail
Quantify Gene Expression via qPCR
GPC1, HAS1, HAS2, HAS3, EXT1, EXT2, DEFB1, TJP1, and CLDN3 expression was quantified with SYBR® Green Real time PCR Master Mix (Sigma). Reactions were carried out as previously described (5 (link)). The 2−ΔCt method was used to calculate relative mRNA expression normalized to the housekeeping gene GAPDH. The 2−ΔΔCt method was used for calculating fold change in gene expression levels versus untreated controls. All the primers were synthesized by Sigma-Aldrich as previously described in
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Defb1
Ext1
Ext2
Gapdh
Gene expression
Has2
Housekeeping gene
Mrna
Primers
Real time pcr
Sybr green
Tjp1
Corresponding Organization : University Medical Center Groningen
Top 5 similar protocols
Variable analysis
independent variables
- Treatment
- GPC1 expression
- HAS1 expression
- HAS2 expression
- HAS3 expression
- EXT1 expression
- EXT2 expression
- DEFB1 expression
- TJP1 expression
- CLDN3 expression
- GAPDH (housekeeping gene)
- Untreated controls
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