PMA (Propidium Monoazide) PMA 染料 (微生物 v-PCR) | Biotium貨號40019/40013

PMA（propidium monoazide, 單疊氮化丙錠）是一種光反應性 DNA 結合染料(photo-reactive DNA-binding dye )，用於細菌(bacteria)、病毒(viruses)和真菌( fungi)等微生物 PCR (viability PCR, v-PCR)。 PMA 可作為 1 mg 凍乾固體(貨號40013)或 20 mM 水溶液(貨號40019)提供。也可以嘗試 PMAxx™ (40069)，它是 PMA 的卓越替代品。

 v-PCR 的基本機制

PMAxx™ 和 PMA 是對 DNA 具有高親和力的光反應染料。染料嵌入 dsDNA 並在暴露於強烈的可見光下形成共價鍵(covalent linkage)。該反應通過在純化過程中去除修飾的 DNA 和抑制 DNA 聚合酶的模板擴增的組合來抑制修飾的 DNA 模板的 PCR 擴增。因為染料是細胞膜不可滲透的，所以當用染料處理含有活細菌和死細菌的樣品時，只有細胞膜受損的死細菌才容易受到 DNA 修飾。在 qPCR 反應中，死細胞 DNA 將顯示延遲擴增和比活細胞更高的 Ct。

PMAxx™ 或 PMA 等活性 PCR 染料不透膜，這使它們具有死細胞特異性。一旦進入死細胞，它們就會與 DNA 結合。暴露在強烈的可見光下會使染料具有反應性，並使它們共價結合到 DNA 上。這種 DNA 修飾可防止後續 PCR 反應中的擴增。Viability PCR dyes like PMAxx™ or PMA are membrane-impermeant, which makes them dead cell specific. Once inside of a dead cell, they bind to DNA. Exposure to intense visible light renders the dyes reactive, and causes them to covalently attach to the DNA. This DNA modification prevents amplification in subsequent PCR reactions.

The basic v-PCR workflow involves the following 5 steps: dye addition, incubation, light exposure, DNA extraction, and qPCR.

• Selectively detect viable cells using qPCR
• Validated in hundreds of publications
• Dead cell specific dye, binds to DNA
• Covalently attaches to DNA after photoactivation
• Available as 1 mg lyophilized solid or 20 mM solution in water
• λ_Abs = 464 nm (before photolysis); λ_Abs/λ_Em= ~510/~610 nm (with DNA/RNA, after photolysis)

To learn more about the advantages of determining microbial or cell viability using viability PCR, visit the Viability PCR Technology Page.

自 Biotium 首次開發 PMA 染料以來，已有數百篇關於該染料在多種樣品類型中的應用的出版物，包括數十種細菌菌株、生物膜、酵母、真菌、病毒和真核細胞。它已用於食品和水安全以及環境測試等應用，並已與 qPCR、NextGen 測序 (NGS)、Sanger 測序和環介導等溫擴增 (LAMP) 結合使用。PMAxx 技術受已授予和/或正在申請的美國和國際專利保護。

About PMA

PMA dye is a DNA modifier invented by scientists at Biotium. It is a photo-reactive dye that binds to DNA with high affinity. Upon photolysis with visible light, PMA dye becomes covalently attached to DNA. This modified DNA cannot be amplified by PCR. The dye is designed to be cell membrane-impermeable. Thus, in a population of live and dead cells, only dead cells are susceptible to DNA modification due to compromised cell membranes. This unique feature of PMA dye makes it highly useful in selective detection of live bacteria by qPCR.

Since Biotium first developed PMA dye, there have been hundreds of publications on the use of the dye in many sample types including dozens of bacterial strains, biofilms, yeast, fungi, viruses, and eukaryotic cells. It has been used in such applications as food and water safety and environmental testing, and has been used in conjunction with qPCR, NextGen Sequencing (NGS), Sanger sequencing, and Loop-mediated Isothermal Amplification (LAMP).