I found error(s) in long desalt oligos after cloning, why?
All oligos prepared by Sigma Life Science are analysed by mass spectrometry as part of quality control. The smallest mass difference in any single base-substitution is 9 Da (A for T). Our MS instruments routinely achieve 0.05% precision, which provides a mass difference of 3.25 Da for a 20mer primer. This is well under the 9 Da difference associated with a worst-case base switch. Related to answer an above, the smallest mass weight of any single base (insertion or deletion) is 304 Da (T). Mass differences of this magnitude are well above the 0.05% mass accuracy and are readily detected. Since 100% of oligos are analysed by mass spectrometry this should provide our customers with 100% assurance that insertions/deletions/substitutions will be caught within our QC systems and rejected. As DNA synthesis is a step-wise manufacturing procedure with a 99.4% coupling efficiency, there are fractional amounts of species where the normal capping step is incomplete. For most applications, these pp
Related Questions
- I used oligos for cloning, sequenced a clone and found a mutation within the oligo sequence. Can mutations happen in synthetic oligos?
- Why do I get a message "Unable to retrieve account sessions information. Error: "s" table structure cannot be recognized"?
- How long about ezpopsy clothes delivery en France ?
