ZymoBIOMICS® Spike-in Control Detection Assays
For real-time PCR detection of the ZymoBIOMICS® Spike-in Control I (D6320) and ZymoBIOMICS® Spike-in Control II (D6321)
Ensure accurate and reliable results using the ZymoBIOMICS® Spike-in Controls and Detection Assays. When used as exogenous internal controls, the ZymoBIOMICS® Spike-in Controls I & II can aid in monitoring microbial extraction efficiency and quality of samples processed through DNA analysis workflows, including PCR and next-generation sequencing (NGS).
The ZymoBIOMICS® Spike-in Controls I & II are composed of quantified bacteria alien to most microbiomes. When spiked into a sample, these bacteria are easily distinguished from native microflora using PCR or NGS. The accompanying ZymoBIOMICS® Spike-in Control Detection Assays use multiplex real-time PCR to specifically detect and differentiate the microorganisms in these controls from within a variety of sample types, including feces, soil, wastewater, swab collections, and more. Together, the ZymoBIOMICS® Spike-in Controls and Detection Assays are valuable tools for workflow optimization and evaluation, even when working with challenging sample types.
Reliable Detection in Samples with Varying Biomass
ZymoBIOMICS® Spike-in Control I
ZymoBIOMICS® Spike-in Control II
Figure 1. ZymoBIOMICS® Spike-in Control Detection Assays were used to assess spike-in recovery across samples having high (fecal) to low (male urine) biomass. Samples were collected in DNA/RNA Shield or Urine Conditioning Buffer (D3061) and extracted using the ZymoBIOMICS® DNA Miniprep Kit (D4300). Bacterial biomass of each sample was determined by detecting total 16S rDNA using the Femto Bacterial DNA Kit (E2006). Real-time PCR was performed on the Bio-Rad CFX96 Touch Real-Time PCR Detection System.
ZymoBIOMICS® Spike-in Control Detection Assays Enable:
Monitoring of extraction efficiency using real time PCR
Easy detection & differentiation of spike-in organisms from native microflora
Multiplexed detection of spike-in microbes in one easy set-up reaction
Identify Microbial Lysis Bias
Figure 2. The ZymoBIOMICS® Spike-in Control I Detection Assay was used to assess lysis bias between two different extraction conditions. Lysis bias was determined by comparing the amplification the spike-in bacteria. Incomplete or biased lysis was observed in one extraction condition, where the tough-to-lyse microbe, Allobacillus halotolerans, amplified over 1.8 cycles later than the easy-to-lyse microbe, Imtechella halotolerans. Real-time PCR was performed on the Bio-Rad CFX96 Touch Real-Time PCRDetection System.
Technical Specifications
| ZymoBIOMICS® Spike-in Control I Detection Assay | |
|---|---|
| Use With: | ZymoBIOMICS® Spike-in Control I (D6320) |
| Recommended Sample Types: | High Biomass (e.g., Fecal, Soil, Wastewater) |
| Multiplex PCR Dyes: |
FAM (ex. 495 nm, em. 520 nm) HEX (ex. 535 nm, em. 556nm) |
| ZymoBIOMICS® Spike-in Control II Detection Assay | |
|---|---|
| Use With: | ZymoBIOMICS® Spike-in Control II (D6321) |
| Recommended Sample Types: | Low Biomass (e.g., Urine; Nasal, Oral, Vaginal Swabs) |
| Multiplex PCR Dyes: |
FAM (ex. 495 nm, em. 520 nm) HEX (ex. 535 nm, em. 556nm) Quasar 670 (ex. 647nm, em. 670nm) |