Gene Expression Analysis by qPCR

Total RNA was extracted using the PureLink RNA Mini Kit (Thermo Fisher), followed by cDNA synthesis using M-MuLV Reverse Transcriptase (New England Biolabs, Ipswich, MA). Quantitative PCR was then performed using TaqMan Fast Advanced Master Mix or PowerUp SYBR Green Master Mix (Thermo Fisher). Glyceraldehyde 3-phosphate dehydrogenase (GADPH) or 18S rRNA served as internal controls (8 (link), 25 (link)). Primer sequences are listed in Table S1.

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Xiao Y., Lian H., Zhong X.S., Krishnachaitanya S.S., Cong Y., Dashwood R.H., Savidge T.C., Powell D.W., Liu X, & Li Q. (2022). Matrix metalloproteinase 7 contributes to intestinal barrier dysfunction by degrading tight junction protein Claudin-7. Frontiers in Immunology, 13, 1020902.

Publication 2022

PureLink RNA Mini Kit M-MuLV Reverse Transcriptase TaqMan Fast Advanced Master Mix PowerUp SYBR Green Master Mix

18s rrna Cdna Glyceraldehyde 3 phosphate dehydrogenase M mulv Primer Reverse transcriptase Sybr green Synthesis

Corresponding Organization : The University of Texas Medical Branch at Galveston

Other organizations : Binzhou Medical University, Texas Children's Hospital, Baylor College of Medicine

Top 5 similar protocols

Variable analysis

independent variables

Reverse Transcriptase (M-MuLV)

dependent variables

Quantitative PCR

control variables

Glyceraldehyde 3-phosphate dehydrogenase (GADPH)
18S rRNA

positive controls

None specified

negative controls

None specified

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