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Applied Biosystems™ Eukaryotic 18S rRNA Endogenous Control (FAM™/MGB probe, non-primer limited)
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Catalog No. 4333760T
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Catalog No. 4333760T Supplier Applied Biosystems™ Supplier No. 4333760T
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Description

Eukaryotic 18S rRNA Endogenous Control (FAM™/MGB probe, non-primer limited)

The Applied Biosystems™ Eukaryotic 18S rRNA Endogenous Control (FAM™ MGB Probe, Non-Primer Limited) is intended as an endogenous control. It allows relative gene expression quantification in cDNA samples when used with other gene expression assays. Probe is labeled with 6FAM™ dye - MGB and the primers are not limited. Can be used for singleplex PCR reactions. Endogenous control is to be used with Inventoried and Non-Inventoried TaqMan™ Gene Expression Assays, Custom TaqMan™ Gene Expression Assays, and⁄or Custom TaqMan™ Primers and Probes.

Assay Details:
Gene Symbol: 18S
RefSeq: X03205.1
Probe Exon Location: NA
Amplicon Size: 187
Corresponding TaqMan Assay ID: Hs99999901_s1

TaqMan™ Endogenous Controls
Eliminate months of assay design, formulation, and testing by using TaqMan™ Endogenous Controls as your controls to quantify gene expression. This convenient collection of pre-designed probe and primer sets enables you to normalize the amount of sample RNA or DNA in a reaction.

Complete Solution for Quantitative Gene Expression
Having a hard time deciding what controls to use to quantify gene expression—even with detailed information on biological systems? Now, with TaqMan™ Endogenous Controls, you can avoid all the trial-and-error of selecting controls for most common human, mouse, rat, and eukaryotic genes.

Simple to Use
All components of the TaqMan™ Endogenous Controls are QC tested, formulated into a single 20X mix, and functionally tested. The controls are not only simple to use, but they are also fully compatible with universal conditions for two-step RT-PCR. Just add TaqMan™ Universal PCR Master Mix (with or without AmpErase™ UNG) and your cDNA sample to generate sensitive, reproducible, and truly quantitative gene expression data on Applied Biosystems instruments including the Applied Biosystems 7900HT, 7300, 7500 Real-Time PCR Systems, and the 7000 and 7700 Sequence Detection Systems.

Flexible Offering
We build each endogenous control using our proven 5' nuclease chemistry. For maximum flexibility, you can choose between two different reporter dyes and two quenchers:
  • A FAM™ dye-labeled TaqMan™ MGB probe (250nM, final concentration) and two unlabeled PCR primers (900nM each)
  • A VIC™ dye-labeled TaqMan™ MGB probe (250nM, final concentration) and two unlabeled PCR primers (150nM each y primer limited)
  • A VIC™ dye-labeled TAMRA™ probe (250nM, final concentration) and two unlabeled PCR primers (150nM each y primer limited)

    Choosing the Right Endogenous Control
    Endogenous controls can normalize the expression levels of target genes by correcting differences in the amount of cDNA that is loaded into PCR reaction wells. For best results, verify that the endogenous control is consistently expressed in the sample set to be tested. Endogenous control expression must be uniform across all samples in the study. For multiplexing, ensure that the gene expression level of the endogenous control is greater than that of the target.

    Multiplex vs. Singleplex PCR
    All TaqMan™ Endogenous Controls that contain probes labeled with the VIC™ reporter dye are primer limited. This allows multiplexing of TaqMan™ Endogenous Controls with target gene expression assays, provided that the control gene is more abundantly expressed than the target gene. All TaqMan™ Endogenous Controls that contain probes labeled with the FAM™ reporter dye are not primer limited and are not intended for multiplexing .

    For Research Use Only. Not for use in diagnostic procedures.

    • Specifications

    Concentration 20X
    Accession Number X03205.1
    Amplicon Size 187
    Content And Storage Control Primer-Probe Set(s)
    Detection Method Primer-probe
    Form Liquid
    Format Tube
    PCR Method 2-step RT-qPCR
    Gene Symbol 18S
    Label or Dye FAM
    Product Line FAM, TaqMan
    Quantity 125 Reactions
    Shipping Condition Room Temperature
    Species Eukaryotic, Rat, Mouse, Human
    Target 18S rRNA
    For Use With (Application) Gene Expression
    No. of Reactions 125 Reactions
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    Frequently Asked Questions (FAQs)

    I am working on both mouse and human samples for gene expression studies. Do you have one endogenous control that I can use with both species?

    The 18S rRNA is highly conserved among eukaryotes. We have several configurations of assays for this gene: P/N 4331182, Assay ID: Hs99999901_s1; PN 4319413E, 4310893E, 4333760T, 4333760F, 4308329. You can use any of these for both human and mouse samples, as well as samples from any other eukaryotic species.
    Are there any TaqMan Pre-Developed Assay Reagents (PDARs) that have not been made obsolete?

    The PDARs for several genes were simply renamed as the TaqMan Endogenous Controls and are available in the different tube sizes in FAM/MGB and VIC/MGB dye and quencher combinations. Those genes are: 18S rRNA; GAPDH; ACTB; TBP; PGK1; GUSB; PPIA; B2M; RPLP0; and TFRC.

    Will TaqMan Gene Expression Assays work similarly to PDARs?

    The TaqMan Gene Expression Assays that were created as replacements for the PDARs were designed with exactly the same sequence as the PDAR. Additionally, the assays are formulated with the same primer and probe concentration in a 20X solution and are available in 250 reaction size. There is virtually no difference between the PDAR and its replacement assay.
    I am trying to recreate a gene expression experiment that used the discontinued PDAR (Pre-Developed Assay Reagents) products. Can I find a TaqMan Gene Expression Assay with the same sequence?

    Yes, we have maintained a TaqMan Gene Expression Assay ID for each PDAR that was available in the past. This assay uses the exact same sequence as the PDAR, and is denoted by a series of five “9's” in the assay ID (example: Hs99999070_m1 is the old PDAR for the gene VEGF).
    What can I use as an alternative to TaqMan Pre-Developed Assay Reagents (PDARs)?

    The TaqMan Gene Expression Assays are the improved alternative for the PDARs. With over 400,000 pre-designed assays for human, mouse, and rat designed over most exon-exon junctions of transcripts in the public RefSeq database, chances are good that you will find the right assay for your experiment. Many assays are available for each gene because assays are designed for each transcript. Therefore, TaqMan Gene Expression Assays will enable you to detect different expression levels for splice variants of genes.
    What are the key features of a TaqMan MGB probe?

    TaqMan MGB probes have a nonfluorescent quencher at the 3´ end and a minor groove binder at the 3´ end that increases the melting temperature (Tm) of probe, allowing the use of shorter probes.  For more information on MGB probes, please consult the following manual: "Primer Express v1.5 and TaqMan MGB Probes for Allelic Discrimination: All PCR Instruments: User Bulletin." You can find a copy on our website by entering this title in the main search box.

    When using the TaqMan Gene Signature Plates, should I mix the reactions after adding the master mix and template to the wells containing lyophilized primers and probe?

    No, but it is recommended that the plate be centrifuged to draw down the liquid to the bottom of the wells.

    What are your recommendations for the best way to dilute and store my TaqMan probe?

    The probe should be diluted in TE buffer [10 mM Tris-HCl, pH 8.0 (at 25°C), 1 mM EDTA]. We recommend storing your probes, both stocks and working solutions, at -20°C in aliquots. This is to cut down on the number of times the probe is subject to freeze-thaws. For detailed information on diluting primers and probes, please refer to the tutorial entitled "Reconstituting and Diluting Primers and TaqMan Probes". You can find it on our website by entering this title in the main search field.

    For Research Use Only. Not for use in diagnostic procedures.