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Features
- Innovative hydrophobic, low-viscosity PCR encapsulation barrier
- Ensures high success rates in precision applications such as real-time PCR
- Eliminates the need for tube caps, accelerating reaction setup
Product Details
Vapor-Lock is an innovative hydrophobic, low-viscosity PCR encapsulation barrier. As little as 10 µl easy-to-pipet Vapor-Lock creates an effective barrier. Vapor-Lock enables reaction overlays that prevent evaporation and eliminate the need for tube caps. Unlike mineral oil, Vapor-Lock does not contaminate or inhibit PCR.
Performance
Use of Vapor-Lock for reaction overlay eliminates the need to cap tubes. In experiments comparing CT values, similar results were obtained from reactions in capped tubes compared to reactions overlaid with Vapor-Lock in uncapped tubes (see figure " Vapor-Lock eliminates need for tube caps").
Small reaction volumes (e.g., reaction volumes in a 384-well plate) typically exhibit poor reproducibility and are extremely difficult to run successfully on many real-time instruments. However, the Rotor-Gene Q cycler constantly centrifuges samples during a run, keeping the reaction at the bottom of the tube aligned with the optical pathway (see figure " Principle of Vapor-Lock"). When working with small reaction volumes, it is important to avoid any evaporation. Vapor-Lock prevents evaporation and has been succesfully used to ensure successful amplification for reaction volumes as low as 5 µl (see figure " Successful amplification using small reaction volumes").
See figures
Principle
Evaporation can occur during thermal cycling, resulting in changes in the reaction volume and the concentration of each reaction component. Vapor-Lock prevents evaporation, ensuring high success rates in precision applications such as quantitative real-time PCR and high-resolution melting (HRM). Vapor-Lock looks and behaves like aqueous buffer. It sits above the aqueous phase, forming a layer on top of the aqueous sample (see figure " Principle of Vapor-Lock"). Vapor-Lock has low viscosity and low specific gravity, allowing aqueous droplets over approximately 1 µl to drop easily through the Vapor-Lock layer to the aqueous sample underneath.
Long reaction setup times, which can occur when working with large numbers of samples in strip tubes or multiwell plates, increase the risk of sample evaporation which can compromise the success of an experiment. Vapor-Lock completely eliminates the need for caps which accelerates reaction setup. Vapor-Lock overlays the samples, preventing evaporation and increasing the reliability of results.
In contrast to mineral oil, Vapor-Lock does not contaminate or inhibit PCR. The background fluorescence of Vapor-Lock is less than one fluorescence unit (see figure " Minimal autofluorescence of Vapor-Lock").
Vapor-Lock is fully compatible with the QIAgility instrument for high-precision, automated reaction setup. It is also highly suited for use with the Rotor-Gene Q cycler.
| Feature | Details |
|---|---|
| Physical properties | PCR-safe; no known contaminants, including DNA, RNA, enzymes, metals, salts or other trace elements |
| Purity | Synthetic, high-molecular-weight polymer; hydrophobic, colorless, odorless, nonhazardous |
| Autofluorescence | None detected spanning ultraviolet to infra red (excitation 36–680 nm; emission 460–712 nm) |
| Viscosity | ≥5 centistoke |
| Vapor pressure | <1 mm Hg |
| Boiling point | >120°C |
| Flash point | <125°C |
| Solubility | <100 ppb |
See figures
Procedure
Vapor-Lock is compatible with both manual and automated reaction setup. We recommend adding Vapor-Lock to reaction tubes prior to any reaction components. Gentle centrifugation during a normal Rotor-Gene Q run ensures full partitioning of the Vapor-Lock and aqueous reaction components into 2 layers (i.e., Vapor-Lock on top and reaction mix underneath). If not using a Rotor-Gene Q cycler, we recommend briefly centrifuging samples prior to cycling.
Applications
Vapor-Lock is highly suited for use in PCR and HRM applications, including:
- Gene expression analysis
- Genotyping
- Mutation scanning
Supporting data and figures
Successful amplification using small reaction volumes.
Human genomic DNA was used as template for amplification of the human bcl-2 oncogene. The total reaction volume was 5 µl. Reactions were performed in uncapped 0.1 ml Strip Tubes with (red) and without (blue) a 10 µl overlay of Vapor-Lock. Five replicate reactions were performed. Data was acquired on the green channel. Vapor-Lock overlay was essential for amplification success of the reactions. Only samples using the Vapor-Lock overlay resulted in detectable amplification (mean CT = 26.23, SD = 0.10).
Resources
Kit Handbooks (1)
Certificates of Analysis (1)
FAQ
If Vapor-Lock is used to overlay PCR reactions, which volume should be entered as reaction volume in the Rotor-Gene Q Software?
Can Vapor-Lock be used with samples processed on the QIAgility?