Plant Molecular Biology

Control Primers for Arabidopsis

Three primer sets designed for use as internal controls in PCR-based Arabidopsis gene expression analysis. All three primer sets have been functionally validated and optimized by RT-PCR analysis against major Arabidopsis tissues for constitutive expression and specific amplification (see Figures 1 & 2). Further confirmation of those primers, as internal control for data normalization, was conducted in quantitative RT-PCR reactions for pathogen-inducible Arabidopsis genes.
These control primers provide Arabidopsis researchers a simple solution for selecting internal control(s) in gene expression analysis:
  • Sensitive, efficient and cost-effective in a ready-to-use format.
  • Represent different expression levels to match target expression level.
  • Choice of three housekeeping genes to meet customers' specific needs.

Each set contains forward and reverse primers for housekeeping genes:
Product Code Product Name
C3115 Control Primer Set for Arabidopsis CBP20 Gene
Gene Locus: At5g44200
Amplicon Size: 409 bp
C3615 Control Primer Set for Arabidopsis Actin-2 Gene
Gene Locus: At3g18780
Amplicon Size: 257 bp
C3240 Control Primer Set for Arabidopsis UBC Gene
Gene Locus: At5g25760
Amplicon Size: 217 bp
Each primer set (forward and reverse) is suitable for 100 PCR/RT-PCR reactions (for 50 µL reaction volume).

Data Analysis
Figure 1. Gel image of end-point Q-RT-PCR amplification of three housekeeping genes in five Arabidopsis tissues.
Lanes 1: Leaf
Lanes 2: Root
Lanes 3: Stem
Lanes 4: Flower
Lanes 5: Silique
  Total RNA was isolated from five tested tissues and treated with DNase I to reduce genomic DNA background. RT-PCR was performed against control primers with 160 ng RNA for each reaction. Amplicons from each RT-PCR reaction were resolved on 2% agarose gel.
Figure 2. Quantitative comparison of housekeeping gene expression in different tissues of Arabidopsis.
Panel A. Experimental setup and CT value
Panel B. Real-Time Q-RT-PCR with control primers
Well# Dye Type Label CT
A1 SBG1 Leaf CBP20 28.015
A2 SBG1 Root CBP20 29.078
A3 SBG1 Stem CBP20 28.199
A4 SBG1 Flower CBP20 26.846
A5 SBG1 Silique CBP20 30.692
Well# Dye Type Label CT
B1 SBG1 Leaf Actin-2 24.921
B2 SBG1 Root Actin-2 23.184
B3 SBG1 Stem Actin-2 24.811
B4 SBG1 Flower Actin-2 24.533
B5 SBG1 Silique Actin-2 26.753
Well# Dye Type Label CT
C1 SBG1 Leaf Ubiquitin 29.631
C2 SBG1 Root Ubiquitin 29.866
C3 SBG1 Stem Ubiquitin 30.801
C4 SBG1 Flower Ubiquitin 28.283
C5 SBG1 Silique Ubiquitin 29.276

Related Products
Product Code Product Description
PR0100 Primer Library for Arabidopsis Pathogen-Inducible Genes 1 Set
QR0100 SYBR® Green Quantitative RT-PCR Kit 1 Kit
J3520 JumpStart™ RED HT RT PCR Kit 40 Reactions
200 Reactions
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