---
title: qPCR assay Q&A (Clone)
description: In a qPCR assay using probe, would non-specific amplification be possible occur in a situation of very high concentration of non-target DNA?
image: https://cdn2.hubspot.net/hubfs/3796365/qa-image.png
---

| × |  |   |
| --- | --- | --- |

|  | powered by | ![](https://www.google.com/cse/static/images/1x/googlelogo_grey_46x15dp.png) |
| --- | --- | --- |

Custom Search

|  | Sort by: Relevance Relevance Date |
| --- | --- |

| Web       |  |
| --- | --- |

[![vast.png](https://www.idexxbioanalytics.com/hs-fs/hubfs/IDXBIO_No_TAGLINE_Pantone.png?width=116&name=IDXBIO_No_TAGLINE_Pantone.png "vast.png")](https://www.idexxbioresearch.com/)

![Discovery-Center-Q&A.png](https://www.idexxbioanalytics.com/hs-fs/hubfs/Discovery-Center-Q%26A.png?width=2100&name=Discovery-Center-Q%26A.png "Discovery-Center-Q&A.png")

**Q ****In a qPCR assay using probe, would non-specific amplification be possible occur in a situation of very high concentration of non-target DNA? **

During specificity test of a diagnostic, when it challenged against plasmid DNA from a non-target species at a very high concentrations of DNA non-target (10E8 copies/ul) there was amplification. However, at lower concentrations this did not occur. Non-specific amplification may have occurred due to stochastic events, ie, since all the components necessary for amplification to occur occur (except for the target DNA), the physical interaction between the molecules could have favored non-specific amplification, considering this extreme situation of High concentration of non-target DNA? 

**A ****In short, yes, it can happen. **

In real-time PCR assays that are designed to be very sensitive you can sometimes have amplification of a nonspecific target. There are several possibilities. Real-time PCR assays, although often more specific than conventional PCR assays because of additional specificity provided by the probe. However, some assays will tolerate one or more mismatches in the probe and/or primer sequence. One possibity, then is amplification of a phylogenetically related organism that has a similar sequence. 

In a qPCR assay using probe, would non-specific amplification be possible occur in a situation of very high concentration of non-target DNA? 

### Posts by Topic

- [How To Video (4)](https://www.idexxbioanalytics.com/blog/topic/how-to-video)
- [Questions & Answer (3)](https://www.idexxbioanalytics.com/blog/topic/questions-answer)
- [What's New (2)](https://www.idexxbioanalytics.com/blog/topic/whats-new)

### Recent Posts

![IBA-logo-stacked_white_Left-just-2](https://www.idexxbioanalytics.com/hs-fs/hubfs/IBA-logo-stacked_white_Left-just-2.png?width=125&name=IBA-logo-stacked_white_Left-just-2.png "IBA-logo-stacked_white_Left-just-2")

IDEXX Laboratories, Inc.  
One IDEXX Drive  
Westbrook, Maine 04092  
United States

IDEXX BioAnalytics  
4011 Discovery Drive  
Columbia, MO 65201  
Phone: 800 669 0825   
573 499 5700  
Fax: 573 499 5701

IDEXX BioAnalytics  
2825 KOVR Drive  
West Sacramento, CA 95605  
Phone: 800 544 5205 x2  
Fax: 916 372 2783

IDEXX BioAnalytics  
3 Centennial Drive  
North Grafton, MA 01536  
Phone: 800 544 5205 x2  
Fax: 508 887 7901

© 2017 IDEXX Laboratories, Inc. All rights reserved.