Reagents for the preparation of two oligonucleotides per synthesis (TOPS).
In order to increase the efficiency of use of automated DNA synthesizers (i.e. the number of oligomers prepared per day), we have devised and prepared novel phosphoramidite reagents that contain a linking group which, while stable under the normal synthesis conditions, is cleaved under basic conditions. When one of these linkers is introduced at the desired position in the synthesis of an oligonucleotide, subsequent detritylation enables the synthesis of a second oligonucleotides sequence upon the first. During deprotection of the oligonucleotide with ammonium hydroxide, the chain is cleaved at either side of the points of introduction of the novel reagent, generating two oligonucleotides free in solution. These reagents are of particular use in applications where oligomers are used in pairs (such as PCR, chemical synthesis of genes etc.) and means that an automated synthesis facility can be used more efficiently, without the need for operator intervention, after the working day is over.
Hardy
P M
PM
Cambridge Research Biochemicals, Northwich, Cheshire, UK.
Holland
D
D
Scott
S
S
Garman
A J
AJ
Newton
C R
CR
McLean
M J
MJ
D000577:Amides / Q000737:chemistry; D064753:Ammonium Hydroxide; D001322:Autoanalysis; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006860:Hydrogen Bonding; D006878:Hydroxides; D007202:Indicators and Reagents; D000432:Methanol; D008744:Methylamines; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009841:Oligonucleotides / Q000138:chemical synthesis*; D010756:Phosphoric Acids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D000515:alpha 1-Antitrypsin / Q000235:genetics
D000577:Amides; D006878:Hydroxides; D007202:Indicators and Reagents; D008744:Methylamines; D009841:Oligonucleotides; D010756:Phosphoric Acids; D000515:alpha 1-Antitrypsin; D064753:Ammonium Hydroxide; C011067:phosphoramidic acid; C027451:methylamine; D000432:Methanol