Total RNA from EBV B cells carrying the non-risk or risk TNIP1 H1 haplotype was isolated using Direct-zol RNA MiniPrep Plus kit (Zymo Research, Irvine, CA) according to the manufacturer’s protocol (3 (link)). cDNA synthesis was performed using QuantiTect reverse transcriptase kit (Qiagen, Germantown, MD) as per the manufacturer’s recommendations. Gene expression was measured by real-time PCR analysis using RT2 SYBR Green (Qiagen). Gene expression primers for human ANXA6 (QT00066941), human DCTN4 (QT00038766), human GM2A (QT00071967), human SMIM3 (QT01028090), human TNIP1 (QT00044072), and human GAPDH (PPH00150F) were purchased from Qiagen.
Protocol detail
Transcriptional Analysis of TNIP1 Haplotypes
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B cells
Cdna
Gapdh
Gene expression
Haplotype
Human
Primers
Real time pcr analysis
Reverse transcriptase
Sybr green
Synthesis
Corresponding Organization : University of Oklahoma Health Sciences Center
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Variable analysis
independent variables
- TNIP1 H1 haplotype (non-risk or risk)
- Gene expression of ANXA6
- Gene expression of DCTN4
- Gene expression of GM2A
- Gene expression of SMIM3
- Gene expression of TNIP1
- Total RNA from EBV B cells
- Direct-zol RNA MiniPrep Plus kit (Zymo Research, Irvine, CA)
- QuantiTect reverse transcriptase kit (Qiagen, Germantown, MD)
- RT2 SYBR Green (Qiagen)
- GAPDH gene expression primer (Qiagen)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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