Processing
Frequently asked questions
Our Scientific Applications Support team has assembled a list of frequently asked questions to help you find answers quickly. Filter using one or more categories to focus on specific topics, or use the search bar to perform a text search.
Can I directly synthesize oligos with sticky ends rather than doing a restriction digest?
Yes, you can design your oligonucleotides with the desired overhangs and ligate them directly into a digested vector.
If you order the oligos with the desired overhangs, you may wish to add a 5' Phosphorylation modification to annealed oligos prior to ligating into your digested vector to increase the ligation efficiency.
You can order the oligos with the modification or perform a phosphorylation reaction to your oligos after receiving them. When you digest your vector with a restriction enzyme, the restriction enzyme will leave a 5' phosphorylation on the vector overhangs that will allow the oligo overhangs to be ligated into the vector.
If you order the oligos with the desired overhangs, you may wish to add a 5' Phosphorylation modification to annealed oligos prior to ligating into your digested vector to increase the ligation efficiency.
You can order the oligos with the modification or perform a phosphorylation reaction to your oligos after receiving them. When you digest your vector with a restriction enzyme, the restriction enzyme will leave a 5' phosphorylation on the vector overhangs that will allow the oligo overhangs to be ligated into the vector.