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Macromolecular Crystallographic Unit
About Instrument and its applications-
1. Macromolecular Crystallographic Unit (MCU): MCU facility at IIC, IIT Roorkee was established for sample preparation to 3D structure determination of macromolecules. MCU has facility for cloning, expression, purification, crystallization, X-ray diffraction and three-dimensional structure analysis of proteins and other bio-macromolecules (nucleic acids, viruses and ribosomes). The knowledge of accurate molecular structures is a pre-requisite for rational drug design, and for structure-based functional studies to aid the development of effective therapeutic agents and drugs. Drug molecules can be identified and designed against potential drug targets for various diseases using this facility. Faculty and students can now target a specific protein of a pathogen (bacterial or viral) to maximize a drug’s effectiveness while at the same time minimizing possible side effects. Structure-based drug design is a relatively new technique that dramatically shortens drug development over the classic trial-and-error method of testing drugs. The resulting structural information is used as a powerful research tool in the pharmaceutical, chemical, agriculture, food and biotechnology industries. 
The Unit is equipped with the instruments required for: 
  • Sample preparation for macromolecular crystallography. 
  • X-ray Diffraction and 3D Structure Determination (Data collection, data processing, structure solution and molecular modelling).
2. Isothermal titration calorimetry(ITC): ITC is a biophysical technique used in quantitative studies of a wide variety of biomolecular interactions. It is a powerful tool for label-free measurement of the binding affinity and thermodynamics of biomolecular interactions to understand function and mechanisms at a molecular level. It works by directly measuring the heat that is either released or absorbed during a biomolecular binding event. In macromolecular Crystallography, it is an integral part of the unit to make protein-ligand and protein interaction studies. ITC is considered as the “gold-standard” method for studying molecular interactions and  its highly useful for enhancing the crystallization of complexes. Complex crystallization can be performed successfully if appropriate preliminary characterization of the complex is carried out through ITC.
 
General Information-
Make- Bruker
Model- Kappa APEX-II
Year of Installation- 2008
 
Specification-
  • Sample Size: 1ng plus
  • Internal Precision:   ~10ppm 2SE for samples over 10ng                                                                                                                                                   ~10-20ppm 2SE for samples less than 10ng
  • External precision:   ~10ppm 2SE for stds over 10ng                                                                                               ~20ppm 2SE for stds under 10ng
  • Accuracy: ~ 30 ppm
  • No. of Faraday Collectors:   9
Sample Requirements-
Sample requirements for MCU: A 3D crystal with smallest dimension of crystal with 50 mm.
Sample requirements for ITC:Researchers should bring pure protein and ligand sample to caary out the experiment. Both the cell and syringe sample components should be prepared only in biologically acceptable and instrument compatible buffering systems and should not include any components that make the sample viscous. Sample should not contain any corrosive substances, strong acids, strong detergents or organic solvents. Reducing agents such as DTT cause erratic baseline drift and other artefacts. TCEP is recommended over BME. Avoid if possible or keep ≤ 1 mM if absolutely required.
 
Analytical Charges-
For Charges Information Click Below Link-
 
Procedure for Sample Submission-
For users of IIT Roorkee Only
Internal candidates have to first register themselves at following portal-
Obtain a Token ID after logging in to above portal and then submit application (which can be downloaded from below link) mentioning the token ID, registration no. along with other relevant details to the IIC office.
For Downloading application click here-
 
For users other than IIT Roorkee
The Application (duly forwarded by concerned department HOD/Guide) along with analytical charges (in the form of Demand Draft) mentioning all the relevant parameters related to analysis may be sent to the below mentioned address-
Head
Institute Instrumentation Centre
Indian Institute of Technology Roorkee
Roorkee-247667
 
Mode of Payment:
For users of IIT Roorkee Only
The payment may be made in advance (In Cash) at Institute Instrumentation Centre Office.
 
For users other than IIT Roorkee
The payment should be made in the form of a Demand Draft in favor of “Dean SRIC, IIT Roorkee”. The DD should be sent along with your application and Samples.
NOTE: The DD amount should include service tax as per GOI rules.
 
Coordinator
Prof. in Department of Biotechnology
Office Tel: 01332-286286/286312
 
Contact Person
Prof. in Department of Biotechnology
Office Tel: 01332-286286/286312