One tube containing lyophilized oligo adaptor. The product is supplied as a lyophilized powder. Oligo purity is greater than 98% as determined by denaturing polyacrylamide gel electrophoresis.
Reconstitute oligo adaptor in sterile water preferably at 100 µM concentration. The total nmol supplied is on the tube label and/or specification sheet. For example if the total quantity is 3 nmols then dissolve in 30 µL. Traditionally molecular biology labs reconstitute at 1 µg/µL but this becomes tedious to convert back to pmol ends for ligation purposes.
Adaptors are short oligos that are supplied in an annealed form. Due to the short size the Tm is low and thus even room temperature storage should be avoided. Always keep them on ice when in use.
After reconstitution store at -20oC
The supplied adaptors are short synthetic oligonucleotide pre-annealed duplexes with 5' blunt end. These can be ligated to the DNA template of interest by blunt end ligation or the cohesive ends These have an internal restriction endonuclease site, which is created by ligation to fragments with complementary overhangs. The duplexes have an overhang and a blunt end.
1. Follow ligase provider protocol for ligation.
2. Visit Gene Link web site for Ligation Calculator to determine ratio of insert:vector at the following link
3. Conditions for adaptor ligation is the same as for insertion of DNA fragments into a plasmid vector. The recommended molar ratio of phosphorylated adaptor:dephosphorylated vector is 10-100:1. When using phosphorylated vector, the adaptor:vector molar ratio should be >100:1.