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Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography

Published on: September 29, 2011

Hydroxylapatite chromatography.

A V Broadhurst¹

¹Roche Products Ltd., Welwyn Garden City, Hertfordshire, United Kingdom.

Current Protocols in Protein Science

|April 23, 2008

Summary

Hydroxylapatite, a calcium phosphate form, serves as an effective chromatography matrix for proteins and nucleic acids. Protocols detail both laboratory-prepared low-pressure and commercial HPLC methods for purification.

Area of Science:

Biochemistry
Analytical Chemistry
Chromatography

Background:

Hydroxylapatite (also called hydroxyapatite) is a form of calcium phosphate.
It is a versatile matrix used in chromatography.

Purpose of the Study:

To provide protocols for using hydroxylapatite in chromatography.
To cover methods for both proteins and nucleic acids.

Main Methods:

Standard low-pressure chromatography using a laboratory-prepared hydroxylapatite column for protein mixtures.
High-Performance Liquid Chromatography (HPLC) using a commercially available column, applicable to proteins and nucleic acids.
Alternate protocols include column chromatography with step gradient elution or batch binding and step-gradient elution.

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Main Results:

Hydroxylapatite chromatography effectively separates proteins and nucleic acids.
Both low-pressure and HPLC methods are viable for purification.
Step gradient and batch binding methods offer alternative elution strategies.

Conclusions:

Hydroxylapatite is a valuable and adaptable matrix for protein and nucleic acid chromatography.
Detailed protocols facilitate its application in both standard and advanced chromatographic techniques.