Real-time PCR
中文- Update Date：2025-03-11
- Units：Instrumentation Resource Center
[YangMing] StepOnePlus™ PCR System
- Brand/Model: ABI StepOnePlus™
- Instrument Location: Yang-Ming Campus
- Room 467, Biomedical Engineering Building (L)
- Room 710, Biomedical Building (H)
- Room 615, Library, Information & Research Building (J)
- Room B02, Shou-Ren Building (M)
- Term of Use: Reservation required
- Free of charge for NYCU users. Off-campus users, please contact the center before using it.
- Reservation >>> Instrument Reservation System
- Contact center staff：
- Extension：62382
- Email：ycirc@nycu.edu.tw
Introduction:
Real-time fluorescent quantitative PCR technology was introduced by Applied Biosystems in the United States in 1996. This technology marked a leap from qualitative to quantitative PCR and demonstrated stronger specificity compared to conventional PCR, effectively addressing the issue of PCR contamination. Due to its high degree of automation, real-time PCR technology has been widely utilized in gene expression, virus/pathogen detection experiments, and SNP genotyping assays in the post-genome era.
The unique experimental design process includes Assay Development Guidelines to expedite the identification of conditions and ensure experiment quality. Primer Express is used to design TaqMan Probes and Primers, and reagents for Optimized Quantitative PCR are employed. Standardized PCR conditions and reaction concentrations are utilized, increasing PCR efficiency to 100%.
Features:
Real-time fluorescent quantitative PCR technology was introduced by Applied Biosystems in the United States in 1996. This technology marked a leap from qualitative to quantitative PCR and demonstrated stronger specificity compared to conventional PCR, effectively addressing the issue of PCR contamination. Due to its high degree of automation, real-time PCR technology has been widely utilized in gene expression, virus/pathogen detection experiments, and SNP genotyping assays in the post-genome era.
The unique experimental design process includes Assay Development Guidelines to expedite the identification of conditions and ensure experiment quality. Primer Express is used to design TaqMan Probes and Primers, and reagents for Optimized Quantitative PCR are employed. Standardized PCR conditions and reaction concentrations are utilized, increasing PCR efficiency to 100%.
Features:
- The fast reaction mode only takes 40 minutes, and the standard mode can complete the reaction within 2 hours. Up to 96 reactions can be completed.
- The exclusive Veriflex heating block technology has six independent temperature controls, allowing it to simultaneously perform six different PCR reaction conditions.
- It features a four-color fluorescence detection system, making conducting Multiplex Real-Time PCR experiments easy.
- The software integrates all experimental applications, including:
- Absolute quantification (Standard Curve)
- Relative quantitative (ΔΔ Ct Method & Relative standard Curve)
- Melting Curve Analysis
- SNP genotyping Assay
- Presence/Absence Assay (qualitative interpretation)
- The software interface provides intuitive guidance for setup.
- Please make an online reservation before use. (For detailed instructions, please refer to the reservation rules)
- Users are responsible for promptly filling in the registration book and retrieving their samples after the experiment. The center will not be responsible for any damage to the sample.
- Users are not allowed to perform the following operations on the computers of the center:
- Change programs or parameters or other people's files.
- Externally connect any peripheral devices.
- Copy or install any program on the center's computer (the programs on the center's computers are all original and have copyrights).
- Violators will be prohibited from using it forever. Violators will be punished according to the school rules for any damage caused.
- If you have any questions when using it, please ask the instrument center technicians.
- Restricted to use 0.1ml PCR 96-well plate or tube. If you use other specifications and cause damage to the machine, you will be liable for repair and compensation!
- The PCR machine will automatically switch to sleep mode 4 hours after completing the experiment. The computer will display "Instrument connection was lost," which is normal.
- If the data download from the computer is unsuccessful, please download it from the PCR machine with a flash drive before the following user starts a new experiment to avoid data overwriting.
- Before starting the experiment, please ensure that the data-collection function is activated. Clicking on this icon by mistake will close the data collection (the icon will appear grayed out), and the machine will not detect real-time information during the experiment.
- It is crucial that the person making the reservation is the actual user and not booking on behalf of someone else.
- The earliest available appointment is two weeks in advance. For example, if today is Tuesday, the earliest available reservation would be the following Monday.
- Each user can reserve a maximum of two sessions at a time, with a required 3-hour interval between the sessions. Additionally, each slot cannot exceed 3 hours.
- We understand that unforeseen circumstances can occur, but if you are more than 30 minutes late for the reserved period, it will be considered as giving up the slot. This policy ensures a fair and efficient use of the reservation system for all users.
- The access or return time of accessories should not affect the use of others.
- Please take your belongings with you and do not leave any items behind (especially samples).
Note:
Cancellations must be made 6 hours before the reserved period to avoid causing inconvenience to others. Frequent no-shows without prior cancellation will result in strict limitations on future reservations after investigation.
- Analysis software: StepOnePlus V2.3
- Operation Manual (StepOnePlus Quick Start)
- Operation Manual (StepOnePlus Experiment Setup)
- Operation Manual (StepOnePlus Analysis process)
- Primer Express v3.0 Chinese operation manual (CD can be borrowed from the center)
- Education Training Handouts (Basic / Advanced & Troubleshooting)
- Instructional Video: Part 1 (Play 1:02:03)
- Instructional Video: Part 2 (Play 53:36)
- Real-time PCR Learning Center from the Official website
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