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Analysis of DNA ejection from the phage by Static light scattering (SLS) (CAT#: STEM-MB-0585-WXH)

Introduction

The ejection of DNA from a bacterial virus (i.e., phage) into its host cell is a biologically important example of the translocation of a macromolecular chain along its length through a membrane. The simplest mechanism for this motion is diffusion, but in the case of phage ejection a significant driving force derives from the high degree of stress to which the DNA is subjected in the viral capsid. The translocation is further sped up by the ratcheting and entropic forces associated with proteins that bind to the viral DNA in the host cell cytoplasm.




Principle

Static light scattering is a technique in physical chemistry that measures the intensity of the scattered light to obtain the average molecular weight Mw of a macromolecule like a polymer or a protein in solution. Measurement of the scattering intensity at many angles allows calculation of the root mean square radius, also called the radius of gyration Rg. By measuring the scattering intensity for many samples of various concentrations, the second virial coefficient, A2, can be calculated.

Applications

The main applications of static light scattering is molecular mass determination of macromolecules, such as proteins and polymers, as it is possible to measure the molecular mass of proteins without any assumption about their shape.

Procedure

1. Sample preparation
2. Measurement by SLS instrument
3. Data analysis

Materials

• Right-Angle Light Scattering (RALS) Detector
• Low-Angle Light Scattering (LALS) Detector
• Hybrid RALS/LALS Detector
• Multi-Angle Light Scattering (MALS) Detector
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