Linked Life Data

10976804

Source:http://linkedlifedata.com/resource/pubmed/id/10976804

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2000-12-5
pubmed:abstractText
Purification of target oligodeoxyribonucleotides from failure sequence by-products of synthesis is often required for polymerase chain reaction primers, DNA sequencing and other oligonucleotide applications. We have developed purification protocols based on a reversed-phase mechanism ("trityl on" purification) using a 96-well Oasis HLB extraction plate. The Oasis HLB sorbent combines excellent pH stability with a high loading capacity allowing for single-step purification of 0.2 microM scale synthesis. After sample loading and washing, the oligonucleotide trityl group is cleaved on the plate with 2% trifluoroacetic acid. Target DNA is eluted with acetonitrile-0.36 mM triethylamine acetate, pH 11.3 (10:90, v/v). Typical yield of purified product is 60-95%. Final purity, measured by capillary gel electrophoresis, was found to be 90% or greater. Alternatively, highly pure oligonucleotides can be obtained by a RP-HPLC "trityl off" method using an XTerra C18 column. The use of volatile triethylamine acetate buffer as an ion-pair for RP-HPLC eliminates the need for further desalting.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9673
pubmed:author
pubmed:issnType
Print
pubmed:day
18
pubmed:volume
890
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
167-77
pubmed:dateRevised
2009-1-15
pubmed:meshHeading
pubmed:year
2000
pubmed:articleTitle
Purification of crude DNA oligonucleotides by solid-phase extraction and reversed-phase high-performance liquid chromatography.
pubmed:affiliation
Waters Corporation, Milford, MA 01757, USA. martin_gilar@waters.com
pubmed:publicationType
Journal Article