TOYOBO BIO

One-step Real-Time PCR kit [Probe], [Freeze drying], [High Fedility]

THUNDERBIRD™ Next Probe One-step qRT-PCR 4×Mix (Lyo-Ready Mix) [QRX-101]

가격표
용도
설명
특징
원리
실시예
관련 제품

가격표

THUNDERBIRD™ Next Probe One-step qRT-PCR 4×Mix (Lyo-Ready Mix)

Code No.	포장	저장 온도	설명서 사양	SDS	법규제 ※
Code No.QRX-101	포장1.25mL × 1(250회용/20μL반응)	저장 온도	설명서 사양	SDS	법규제 ※

용도
﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿﻿●  High Efficient One-step qRT-PCR Master Mix (동결건조용)
설명
﻿THUNDERBIRD™ Next Probe One-step qRT-PCR 4×Mix는 고효율 역전사효소인 **ReverTra Ace™**와 Hot Start TTx DNA Polymerase를 기반으로 한 One-step qRT-PCR용 시약입니다.
이 두 효소와 최적화된 버퍼는 소량의 RNA도 빠르고 민감하게 검출 및 정량할 수 있게 해줍니다.
이 믹스는 글리세롤을 포함하지 않으며, 동결건조 시 별도의 첨가제가 필요하지 않습니다.
동결건조 후에도 실온에서 안정적으로 보관 및 아이스팩 없이 운송이 가능하며, 샘플과 물만 추가하면 바로 PCR을 수행할 수 있습니다.
또한, 동결건조가 PCR 성능에 영향을 주지 않습니다.
특징
● 동결건조 가능
동결건조에 필요한 첨가제가 포함되어 있어 별도 구성 없이 동결건조 시약으로 제조가 가능합니다.

● 신속하고 높은 민감도Fast cycling mode가 가능하며, 미량의 RNA도 빠르고 민감하게 검출이 가능합니다.
● 간편한 사용법프라이머, 프로브, 샘플만 추가하면 바로 반응이 시작되므로, 오염 및 피펫팅 오류의 위험을 줄여줍니다.
● 오염물질에 대한 저항성 향상PCR inhibition 존재 하에서 효율적인 증폭이 가능합니다.
● 우수한 멀티플렉스 성능다중 유전자 검출이 가능한 멀티플렉스 반응에 적합합니다.
● dUTP 기반 Carryover 오염 방지 기능 호환dUTP 사용으로 carryover 오염에 의한 위양성(거짓 양성) 결과를 방지할 수 있으며, 별도로 판매되는 UNG와 함께 사용할 수 있습니다.
원리
실시예
1.Detection sensitivity comparison
Using TaqManTM probe, we compared the detection sensitivity and quantitation of mumps virus RNA with other companies' products.
Only this kit was able to detect RNA of less than 10 copies and quantify it over a wide dynamic range. This kit is effective in detecting RNA viruses and mRNAs with low expression levels.




 
2.Detection comparison during fast cycles
Influenza virus RNA was detected using the "Normal" cycle with an elongation time of 45 seconds and the "Fast" cycle with an elongation time of 10 seconds. Analysis was performed using Applied BiosystemsTM StepOnePlusTM.
As a result, efficient amplification was possible even with a fast cycle of 10 s elongation time.

 
3. Comparison of the detection sensitivitiesfor different viruses
The detection sensitivity of five types of viral RNA were cmpared using TaqManTM probes.
Applied BiosystemsTM StepOnePlusTM was used for the analysis.
This product was able to detect all targets with a sensitivity of 10 copies or less.
It is possible to detect various RNAs with high sensitivity, regardless of the primer/probe sequence.

 
4. Tolerant of contaminant
Biological samples (nasal swabs, saliva, and plasma) were added to the reaction to detect 50 copies of influenza virus RNA and evaluate resistance to PCR inhibitors. Bio-Rad CFX96 Touch Deep Well was used for the analysis.
While other company reagents caused poor amplification when PCR inhibitors were added, this reagent showed minimal effect on Ct even when biological samples were added.

 
5.Comparison between singleplex and multiplex methods
Single-and multiplex detections were compared using TaqManTM probes labeled with four different fluorescent dyes. HeLa S3 RNA (10x dilutions of 500 ng to 0.05 ng [5 steps]) was used as a template.
The results revealed that single and multiplex detection showed comparable PCR efficiencies and that multiplex detection did not affect the reaction.

 
6. Carryover prevention by adding UNG
Assuming that the splashed amplified product was contaminated in the subsequent PCR (carryover contamination), we used the amplified sample (196bp) equivalent to 103, 102 copies as a template and detected the same target under the conditions of UNG addition (+)/(-).
Analysis was performed using Applied BiosystemsTM StepOnePlusTM.
As a result, the template was degraded and no amplification was observed when UNG was added.

 
7. Performance comparison before and after lyophilization
qPCR Mix was prepared using THUNDERBIRDTM Next Probe One-step qRT-PCR 4×Mix, and 1500, 375, 94, 23, and 6 copies of influenza A virus RNA were detected before and after lyophilization, respectively. Analysis was performed using a Bio-Rad CFX96 Touch Deep Well.
Good amplification was obtained in all cases and no freeze-drying effects were observed.

 
기타 실시예