Indian Council of Agricultural Research
(Agricultural Knowledge Management Information System)


ICAR-Central Sheep & Wool Research Institute, Avikanagar, via Jaipur-304501, Rajasthan (www.cswri.res.in)
Click Here to See Management Committee
Instrument Details
Instrument/Facility Name: Real Time PCR with Note Book Computer with primer express software & RQ software
Make                                   : Applied Biosystem
Model                                  : Step One Plus
Specification   :Thermal Cycling System: Peltier-Based System, Capacity: 1 x 96-well plate, 12 x 8-tube strips, 96 x 0.1 ml tubes, Run Time: <2 hrs⁄run (Standard Mode), <40 min⁄run (Fast Mode).

Click to ViewThe StepOnePlus™ Real-Time PCR System is a 96-well Real-Time PCR instrument perfect for both first-time and experienced users. The StepOnePlus™ Real-Time PCR System can be setup in a variety of configurations and comes ready to use, out of the box, with intuitive data analysis and instrument control software. Utilizing robust LED based 4-color optical recording, the StepOnePlus™ Real-Time PCR System is designed to deliver precise, quantitative Real-Time PCR results for a variety of genomic research applications. We supply a Dell™ Laptop with the StepOnePlus™ Real-Time PCR System. The StepOnePlus™ Real-Time PCR System draws 38% less energy to process one sample plate (when the instrument is in a heated state), compared to the Bio-Rad CFX96™ Real-Time PCR Detection System. The StepOnePlus™ Real-Time PCR System also has advanced temperature control through use of VeriFlex™ Blocks technology and has a footprint nearly 15% smaller than Bio-Rad CFX96™, which in today’s crowded laboratories helps you use your laboratory space even more efficiently. You also leave a smaller environmental footprint on the Earth. The system measures amplification as it occurs, allowing you to monitor the progress of your experiment cycle by cycle, either on the machine or remotely. Your data is stored on the instrument itself, and can be viewed and stored automatically via remote access, or transferred via email or a USB flash drive. Data can also be conveniently exported as PowerPoint®, Excel®, and graphical image files.

  Working Principles
Click to ViewReal-time PCR is carried out in a thermal cycler with the capacity to illuminate each sample with a beam of light of at least one specified wavelength and detect the fluorescence emitted by the excited fluorophore. The thermal cycler is also able to rapidly heat and chill samples, thereby taking advantage of the physicochemical properties of the nucleic acids and DNA polymerase. The PCR process generally consists of a series of temperature changes that are repeated 25 – 50 times. These cycles normally consist of three stages: the first, at around 95 °C, allows the separation of the nucleic acid's double chain; the second, at a temperature of around 50-60 °C, allows the binding of the primers with the DNA template; the third, at between 68 - 72 °C, facilitates the polymerization carried out by the DNA polymerase. Due to the small size of the fragments the last step is usually omitted in this type of PCR as the enzyme is able to increase their number during the change between the alignment stage and the denaturing stage. In addition, in four step PCR the fluorescence is measured during short temperature phase lasting only a few seconds in each cycle, with a temperature of, for example, 80 °C, in order to reduce the signal caused by the presence of primer dimers when a non-specific dye is used. The temperatures and the timings used for each cycle depend on a wide variety of parameters, such as: the enzyme used to synthesize the DNA, the concentration of divalent ions and deoxyribonucleotides(dNTPs) in the reaction and the bonding temperature of the primers. Real-time PCR technique can be classified by the chemistry used to detect the PCR product, specific or non-specific fluorochromes.

  Applications
  User Instructions
The StepOnePlus™ System supports many Real-Time PCR applications such as: • SNP Genotyping • Gene Expression Analysis • MicroRNA Expression • Protein Expression • Translocation Analysis • Gene Detection • Viral Load Analysis 1. StepOne or StepOnePlus system, including setup, run and analysis. You can perform the following types of experiments on the StepOne and StepOnePlus systems: • Quantitation, including: – Standard curve – Relative standard curve – Comparative CT (ΔΔCT) • Genotyping • Presence/absence. 2. The instrument monitors the progress of the PCR as it occurs.‡ • Data are collected throughout the PCR process. • Reactions are characterized by the point in time during cycling when amplification of a target is first detected. 3. Data are collected at the end of the PCR process. • Reactions are characterized by the quantity of the target accumulated at the end of PCR.§ • The datapoint is the normalized intensity of the reporter dye, or Rn. 4. You can use the following reagent types (chemistries) on the StepOne and StepOnePlus systems: • TaqMan® reagents • SYBR® Green reagents • Other fluorescent-based reagents. SYBR Green reagents include primers and master mixes that contain SYBR® Green dye. You can use SYBR Green reagents for quantitation experiments: • Standard curve • Relative standard curve • Comparative CT (ΔΔCT)

  Contact Us
Contact No. 7568747105
Email Id sganesh413@gmail.com

  Real Time PCR with Note Book Computer with primer express software & RQ software Charges
Procedure Name
Industry
University
National LAB/R&D
Remark
Real-time PCR assay
300/assay
100/assay
100/assay
The charges are for the use of equipment only.

Additional Rs 100/-towards postal charges if result are to be posted
The Demand Draft should be in favour of "ICAR UNIT -CSWRI, Avikanagar payable at SBI, MalpuraTonk Rajasthan"
Letter, DD & Samples send to "Head, Animal Health Division, CSWRI, Avikanagar - 304 501"