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Friday, December 28, 2012

Good Notes on oligonucleotide Yield, Resuspension, and Storage

Oligonucleotides are short stretch of single stranded DNA or RNA molecules used in genetic testing, research and forensic. These are chemically synthesized and has got applications in polymerase chain reaction (PCR),DNA sequencing etc.

Length of oligonucleotide is denoted as '-mer' for example : 18-mer, 22-mer, etc.

Nucleic  acid synthesis columns come in several scales, which are a function of the amount of the 3' nucleoside coupled to the support material. Popular column  sizes Oligonucleotide Synthesis 
range from 40 nmol  to  10umol. Popular support materials  include controlled-pore glass  (CPG) and polystyrene. The CPG columns  are  typically  supplied having pore sizes of either 500 A or 1000 A. The  scale of column used for a synthesis depends on both the quantity and the length of oligonucleotide desired. The greater the amount of 3' nucleoside  attached to the column  support,  the greater the yield of oligo. Polystyrene or large-pore CPG columns  are recommended for oligonucleotides  greater than 50.

oligonucleotides are synthesized at different scales like 0.05umol, 0.2umol, 1umol, etc. depending on the application it is used for.

The number of micromoles of an oligonucleotide synthesized on a column,  therefore, can be calculated by multiplying  the length of the oligonucleotide by the column scale.

A good explanation on calculation of oligonucleotide Yield, Resuspension, and Storage

Integrated DNA Technologies
OligoCalc, Technical guide
Eurofins-MWG Operon,