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 제품안내  분자생물학   PCR Amplification  PCR Kits & Dyes

EvaGreen™ qPCR Dye

SYBR Green I을 대체해서 사용할수 있는 non-toxic, non-mutagenic, non hazardous한 real-time PCR(qPCR)용 dye입니다.

제품 문의하기

▶ EvaGreen Plus

  • 기존 original EvaGreen® Dye 보다 background 가 낮고 signal이 더 높은 qPCR용 dye입니다.
  • 대부분의 qPCR 장비의 excitaion sources에 최적화된 487nm의 excitaion 가지고 있어, background 가 낮기 때문에 더 좋은 결과를 얻으실 수 있습니다.
  • Real-Time PCR 시 Ct (역치 사이클) 값을 더 빠르게 도출해 낼 수 있습니다.
  • digital PCR 또는 isothermal application 방법의 LAMP 증폭 실험과 같이 noise가 많은 기법에서도 높은 signal을 확인 하실 수 있습니다.
  • gel에서 amplification 산물을 직접 눈으로 확인 하실 수 있습니다.

Escitation (left), emission (right) spectra of EvaGreen Plus dye bound to dsDNA in 1M Tris buffer, pH 8.5.

 [original EvaGreen® Dye 와의 비교 데이터]

A comparison of the raw fluorescence signals from qPCR reactions performed with EvaGreen® Plus Dye (Solid, Dark Green) and original EvaGreen® Dye (Dashed, Light Green), two replicates each. EvaGreen® Plus Dye gives an earlier Ct, has a lower background, and higher maximum signal intensity.

A comparison of EvaGreen® Dyes during melt curve analysis. EvaGreen® Plus Dye (Solid, Dark Green) and original EvaGreen® Dye (Dashed, Light Green), two replicates each. The raw fluorescence melt plot (A) reveals that EvaGreen® Plus Dye is brighter when bound to dsDNA and has a lower post-melt background fluorescence. The derivative melt plot (B) displays no significant shift to melt temperatures or DNA disassociation characteristics.

▶ EvaGreen
  • EvaGreen은 기존에 셋팅된 조건의 변화없이 SYBR Green I을 대체하여 사용하실 수 있습니다.
  • "release-on-demand" DNA-결합 기술을 이용하여 PCR inhibition이 거의 없습니다.
  • SYBR® Green I보다 높은 감도로 형광이 매우 밝습니다.
  • 돌연변이를 일으키지 않으며, 세포독성이 없는 제퓸으로 별도의 분리수거 없이 처리가 간편합니다.
  • 보관 또는 PCR 사용시 열, 빛, 산-염기 조건에서도 매우 안정합니다.
  • Fast PCR과 Multiple PCR protocol에 사용 가능합니다.

EvaGreen® dye binds to dsDNA via a "release-on-demand" mechanism.

Figure 1. Comparison among Fast-Plus EvaGreen® master mix from Biotium and two fast SYBR® Green master mixes from two leading companies (company A and company Q) under similar condition. The inset is an enlarged view of the area near the baseline for better viewing the curve patterns of the much weaker signals of the two SYBR-based master mixes. Amplicon: ATPG fragment of human genomic DNA; instrument: ABI 7900 Fast.
Figure 2. Comparison of cell membrane permeability between EvaGreen® dye and SYBR® Green I. HeLa cells were incubated with SYBR® Green I (1.2 μM) or EvaGreen® dye (1.2 μM) at 37 oC. Photographs were taken following incubation for 5 and 30 minutes. SYBR® Green I entered cells rapidly while EvaGreen® dye appeared membrane-impermeable as evident from the absence of cell nuclear staining. Image taken with long photo-exposure time revealed that EvaGreen® dye only associated with cell membranes.
Figure 3. A comparison of the raw fluorescence signal from qPCR reactions performed with two EvaGreen® master mixes (Forget-Me-Not™ EvaGreen® and Fast EvaGreen®) and QuantiNova SYBR® Green. EvaGreen® Dye is less inhibitory than SYBR® green, allowing for a much brighter signal.