Gel electrophoretic analysis of poly(riboadenylic acid)

Area of Science:

• Biochemistry
• Polymer Science
• Molecular Biology

Background:

• Accurate determination of molecular weights and distributions is crucial for understanding polynucleotide and polyelectrolyte behavior.
• Existing methods may have limitations, especially for single-stranded nucleic acids and synthetic polymers.

Purpose of the Study:

• To establish polyacrylamide gel electrophoresis as a method for determining molecular weights and distributions of single-stranded polynucleotides.
• To demonstrate the capability of preparative electrophoresis for obtaining narrow molecular-weight distribution fractions.
• To provide molecular-weight calibration data for poly(rA) and related polymers.

Main Methods:

• Electrophoresis in polyacrylamide gels was employed for molecular weight determination.
• Preparative electrophoresis was used to isolate fractions with narrow molecular-weight distributions from polydisperse samples.
• Sedimentation coefficients were used for molecular-weight calibration.

Main Results:

• Polyacrylamide gel electrophoresis effectively determined molecular weights and distributions for poly(rA) and other single-stranded polynucleotides.
• Narrow molecular-weight distribution fractions were successfully obtained via preparative electrophoresis.
• Molecular-weight calibrations based on sedimentation coefficients were established and shown to differ from those of partially base-paired RNA.

Conclusions:

• Polyacrylamide gel electrophoresis is a viable technique for analyzing molecular characteristics of single-stranded polynucleotides and polyelectrolytes.
• The developed calibration curves allow for rapid molecular-weight determination of poly(rA) and similar structures, including those in messenger RNA.
• The method's utility extends to both synthetic and natural polyelectrolytes.