Distinguish true target negatives from PCR inhibition
Applied Biosystems TaqMan exogenous internal positive control reagents contain a pre-optimized internal positive control (IPC) with predesigned primers and a TaqMan probe. The IPC can be spiked into samples to distinguish true target negatives from PCR inhibition.
TaqMan exogenous internal positive control reagents allow you to simultaneously amplify the IPC and low-copy target DNA in the same tube. Although the target and IPC DNAs may differ in initial copy number, the concentration of the IPC primers in the PCR reaction is limiting, so that the amplification efficiency of the target reaction is not compromised.
Features
TaqMan exogenous IPC reagents are designed to:
Distinguish types of negative results
Distinguish between negative reactions from samples lacking the target sequence vs those containing a PCR inhibitor
Simultaneously amplify two DNAs
By labeling a low-copy target DNA with a FAM probe and the IPC with a VIC probe, the two can be amplified in the same tube
Prevent cross reactivity with target
Permits co-amplification of the IPC and the target sequence without compromising amplification of the target sequence
Avoid amplification of endogenous genes
Maintains expression stability of the endogenous internal control or “housekeeping” gene
How it works
During amplification, the sample and IPC generate reporter fluorescence signals that help to identify unknown samples.
Positive and negative calls are made based on statistical analysis of data from the two dye layers. The statistical analysis sets up threshold values for positive FAM and VIC calls based on the No Template Control (NTC) and the Negative Internal Positive Control (IPC–) baselines.
In this kit, the FAM layer shows the positive (+) and negative (–) calls for the target template and the VIC layer shows the +/– calls for the IPC.
Interpreting negative results with TaqMan exogenous IPC reagents
If the detectable target template (FAM) call is … |
And the detectable IPC (VIC) call is … |
Then the target template is … |
|---|---|---|
+ |
+ or –* |
Present |
– |
+ |
Absent |
– |
– |
No amp |
*In the presence of a strong FAM signal for the target assay, a negative assignment and/or signal can be obtained in the VIC layer. This is a result of the limiting primer concentrations used in the IPC assay.
Ordering information
For Research Use Only. Not for use in diagnostic procedures.