Total RNA was prepared from naïve or differentiated T cells using TriZol reagent (Invitrogen). cDNA was synthesized using Superscript reverse transcriptase and oligo(dT) primers (Invitrogen), and gene expression of beta–Actin (Actb), Grail, Il21, Il4, Gata321 (link),33 –34 (link), Il2 (F-GCGCACCCACTTCAAGCTCCA; R-CTGTGGCCTGCTTGGGCAAGT), Il5 (F-CGCTCACCGAGCTCTGTTG; R-CCAATGCATAGCTGGTGATTTTT), Il13 (F-GCTTATTGAGGAGCTGAGCAACA; R-GGCCAGGTCCACACTCCATA) and Stat6 (F-AGTCACTATAAGCCCGAACAG; R-GCCATTCCAAGATCATAAGGT) was examined with a Bio-Rad iCycler Optical System using iQ SYBR green real-time PCR kit (Bio-Rad Laboratories, Inc.). The data were normalized to Beta-Actin gene.
Protocol detail
Gene Expression Analysis of Differentiated T Cells
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TriZol reagent
Superscript reverse transcriptase
oligo(dT) primers
iCycler Optical System
iQ SYBR green real-time PCR kit
Beta actin
Cdna
Gene
Gene expression
Il13
Il21
Oligo
Primers
Real time pcr
Reverse transcriptase
Stat6
Sybr green
T cells
Trizol
Corresponding Organization :
Other organizations : The University of Texas MD Anderson Cancer Center
Top 5 similar protocols
Variable analysis
independent variables
- Naive T cells
- Differentiated T cells
- Gene expression of beta-Actin (Actb)
- Gene expression of Grail
- Gene expression of Il21
- Gene expression of Il4
- Gene expression of Gata3
- Gene expression of Il2
- Gene expression of Il5
- Gene expression of Il13
- Gene expression of Stat6
- Beta-Actin gene (used for data normalization)
- No positive or negative controls were explicitly mentioned in the input protocol.
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