PrimeTime® qPCR 5' Nuclease Assays
Primer, Probe mix
Real-Time PCR 을 위한 Probe와 2개의 PCR primers를 한 tube에 담아서 제공합니다.
•• 5' reporter dye로 FAM, HEX, TET, Cy5, TAMRA 가능
•• 100% QC
•• 5-7일 안에 사용하기 편한 single tube에 공급
•• PCR을 위한 최적화; ideal for many life science research applications
•• 제품과 함께 sequence information이 있는 Specification sheet 가 제공
•• IDT에서 입증된 Real-Time PCR Design Tool을 통한 디자인
PrimeTime qPCR assays는 세 가지 사이즈로 공급됩니다. primers 와 probe가 premixed 되어 한 tube에 담아서 lyophilized 형태로 공급되며 500 nM의 primer 와 250 nM 의 probe를 반응시 명시된 사용량을 충분히 이용하실 수 있습니다. 주문하시면 약 1주일만에 받아보실 수 있으시 며 다른 회사와 달리 모든 올리고에 대해서 100% QC 가 이루어지게 됩니다.
제품 구성 및 가격
PrimeTime® qPCR 5' Nuclease Assays에 가능한 Reporter 와 Quencher 조합
Pre-designed PrimeTime® qPCR Assay
* 100 % 결과 보장 (positive와 negative control 데이터 제공 시)
* Sequence 제공 (제품과 동봉)
* Human, mouse, rat 세 가지 species에서만 디자인 제공
Pre-designed PrimeTime® qPCR Assay는
아래 링크에서 디자인을 직접 할 수 있습니다.
Overview of 5′ Nuclease Assays
Step 1—The primers and probe hybridize in a sequence-dependent manner to the complementary DNA strand. Because the probe is intact, the fluorophore and quencher are in close proximity and the quencher absorbs fluorescence emitted by the fluorophore.
Step 2—The polymerase extends from the primers and begins DNA synthesis.
Step 3—The polymerase reaches the probe and the exonuclease activity of the polymerase cleaves the hybridized probe. As a result of cleavage, the fluorophore is separated from the quencher and fluoresces.
Step 4—The fluorescence is detected by the real time instrument.
These steps are repeated for each PCR cycle and allow detection of specific products. With intercalating dyes, such as SYBR® Green I, primer-dimers and non-specific products will also contribute to fluorescence. In contrast, the 5’ Nuclease assay is specific and fluorescence will only be detected for the DNA sequence to which the probe and primers hybridize.
Selecting the Correct Reporter Dye and Quencher
The correct reporter dye will depend on the type of instrument you are using and the compatibility of the dye with the instrument. Please see the Instrument Compatibility Table for a list of reporter dyes compatible with common instrumentation.
For multiplexing applications, it is recommended that reporters dyes with the least amount of spectral overlap be selected. For a complete list of IDT’s dyes and quenchers please see the Dye and Quencher Wavelength Figure and the Instrument Compatibility Table
Traditional dark quenchers absorb broadly and do not emit light which allows for the use of multiple reporter dyes with a given quencher. This characteristic allows for expanded options for multiplex assays. Dark quenchers simplify detection which makes them compatible with a broad range of image analysis instruments.
IDT has developed an internal ZEN quencher that enables to production of Double-Quenched Probes which have less background and more signal. The Double-Quenched Probes contain a 5’ FAM fluorophore, a 3’ IBFQ quencher, and an internal ZEN quencher. These Double-Quenched Probes are an improvement over traditional dual-labeled probes and have consistently low background, reduced Cq values, improved precision, and enable the use of longer probes for design in AT-rich regions. For more information download the ZEN Double-Quenched Probe Overview.
IDT supplies commonly used dark quenchers as well as the proprietary dark quenchers, Iowa Black FQ, Iowa Black RQ, and the newly developed internal ZEN quencher. TAMRA is also a quencher option for a FAM reporter dye. Related Information