Since the TaqMan® Rodent GAPDH Control Reagents (Cat. No. 4308313) will also amplify pseudogenes and/or genomic DNA, is there some equation that can be used to correct for genomic DNA contamination?

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Answer

There is no equation for the correction of data for genomic contamination; however, a rule of thumb is that if the Ct difference between your target and the contaminant is 10 or more, you can assume it is not having an effect in your reaction. Please consult the TaqMan® Rodent GAPDH Control Reagents Protocol for more information on these reagents. Enter the catalog number in the main Search field on our web site to view the product detail page and the Protocol.

Answer Id: E1572

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2df02da5a427b102511312bdd5a7e069_FAQ

What is the GenBank accession number of the sequence that was used to design the primers and probe for the TaqMan® Rodent GAPDH Control Reagent Kit (P/N 4308313)?

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Answer

The GenBank accession number used is NM_008084. For more information on this kit, please refer to the TaqMan® Rodent GAPDH kit protocol.

Answer Id: E1811

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3eae6207a26f48a5d2105c15ff641e91_FAQ

What type of control RNA is included in the TaqMan® Rodent GAPDH kit (P/N 4308313)?

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Answer

The RNA included in the TaqMan® Rodent GAPDH kit is total RNA extracted from P388D1 lymphoma mouse cell line.

Answer Id: E1812

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4bf718343dc4549d587f0f8a1a8819f1_FAQ

Are there any TaqMan® Endogenous Controls reagents available that can be used with rat samples?

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Answer

We have TaqMan® Endogenous Controls for 18S Ribosomal RNA and a kit (Cat. No. 4308313) that can be used with rat samples. The Eukaryotic 18S Ribosomal RNA Reagent is in a highly conserved area that allows this TaqMan® Endogenous Control to be used for rat, human and mouse. In addition, we have newer TaqMan® Gene Expression Rat Endogenous Controls, including beta glucuronidase and beta-2 microglobulin. These can be found by searching our Gene Expression Assays and specifying the species of interest as R. norvegicus and the set membership as Endogenous Controls.

Answer Id: E1006

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11390b8013e769146d22fe7fd491eaee_FAQ

Do the human and rodent GAPDH primers and TaqMan® probe sequences manufactured by Applied Biosystems® detect genomic DNA?

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Answer

This depends on the individual probe. Potential reaction with genomic DNA - based on position of primers and/or probe within or across exon junctions - is one of the specifications indicated in the Assay search tool and product support materials. Here are details on the reported performance of a few specific products:

- Taqman® GAPDH Control Reagents, human, 402869, JOE™: Does not detect genomic DNA
- Taqman® Endogenous Control, GAPDH for human, 4326317E, VIC® : Does not detect genomic DNA
- Taqman® Endogenous Control, GAPDH, human, 4310884E, VIC®: Does not detect genomic DNA
- Taqman® Endogenous Control, GAPDH, human, 4333764F, FAM™: Does not detect genomic DNA
- Taqman® Endogenous Control, GAPDH, human, 4333764T, FAM™: Does not detect genomic DNA
- Taqman® Gene Expression Assay, GAPDH, human, Hs99999905_m1/4331182, FAM™: Does not detect genomic DNA

Answer Id: E1517

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61d9eda7bbafd1a3f235723cdc63378f_FAQ

In the TaqMan® Rodent GAPDH Control Reagents kit (P/N 4308313), what rodent is the Control RNA isolated from?

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Answer

The control RNA is total RNA isolated from a mouse lymphoma cell line.

Answer Id: E1128

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470b336d42f2141e9dbb257fcb301958_FAQ

Do your human and rodent GAPDH primers and TaqMan® probe sequences detect genomic DNA?

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Answer

It varies by product, so be sure to check all the available product specifications before purchase. Here are details on a few options:

- The TaqMan® GAPDH Control Reagents (Human), Cat. No. 402869, which is labeled with JOE™ dye, spans two exons therefore will not amplify genomic DNA.

- The Human GAPD (GAPDH) Endogenous Control (VIC®/ TAMRA™ Probe, Primer Limited), Cat. No. 4310884E, is also designed across an exon-exon boundary and will not amplify genomic DNA.

- The probe in the TaqMan® Rodent GAPDH Control Reagents kit, Cat. No. 4308313, labeled with VIC® dye, does not span an intron and therefore may amplify genomic DNA.

Answer Id: E1435

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e0a78d4fcd324aec4d29a45448d7a013_FAQ