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 C_T value

Panel B. Real-Time Q-RT-PCR with control primers

Well#	Dye	Type	Label	C_T
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	C_T
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	C_T
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	Quantity
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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