21 Feb 2012

Using HPLC for Studying DNA and PCR Products

Column liquid chromatography continues to be employed for the separation, purification, along with detection of nucleic acids. Between chromatography methods offered, anion-exchange chromatography continues to be mostly used by the isolation and also purification involving besides oligonucleotides but large double-stranded DNA.

In anion-exchange chromatography, DNA preservation is dependent upon electrostatic interactions between your negatively charged phosphate groups of DNA as well as cationic sites in the chromatographic matrix (begin to see the picture below).

DNA can be eluted on the anion-exchange column by means of altering the ionic strength in the buffer solution. From the reputation of a buffer of improving ionic strength, DNA storage is normally a operate regarding how many negative charges associated with the phosphate groups.

One of the most typical anion-exchange resources is diethylamino ethyl (DEAE)-bonded support, and possesses already been thoroughly utilized to alter initial soft-gel supports, porous microparticular silica, in addition to polymer-based resources. Option of porous microparticular silica in addition to polymer-based supports possesses resulted in improved column resolution as well as more quickly analysis times. Nevertheless, this columns packed with porous particles are used solely along with limited good results from the analysis of large double-stranded DNA fragments.

HPLC columns based on nonporous, small-particle resins are offered, it's very quickly and as well efficient in splitting large double-stranded DNA. The good results of such columns will depend on the truth that nonporous, modest particle measurement material with that they are usually packed provides improved effectiveness in addition to brief analysis time. This can be the consequence of a much faster DNA transfer involving the solid and also liquid column phases than is possible for standard porous support. Moreover, these types of columns offer high DNA recoveries due to absence of pores.

HPLC associated with nucleic acids along with PCR products continues to be productively put to use in post-PCR analyses.

See more: http://bioteachnology.com/pcr/employing-hplc-for-analyzing-dna-and-pcr-products

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