Lightning Terminators provide the increased accuracy, potentially longer read-lengths and faster cycle-times necessary in today’s NGS market.
LaserGen’s innovative Lightning Terminators™ are designed with specialized modifications that increase performance over their conventional reversible terminator counterparts. Our published data indicate that Lightning Terminators are more accurate than natural nucleotides with fast incorporation and cleavage kinetics (Litosh 2011). These properties give the potential for more accurate and faster sequencing reactions.
The difference in our chemistry derives from the location of the functional groups on the nucleotide structure. Reversible terminators generally have two separate functional groups: the terminating group and the label. Lightning Terminators are designed such that the terminating group is attached to a hydroxymethyl handle on the nucleotide base. This attachment leaves the 3’-OH free and results in a more conserved nucleotide structure than conventional reversible terminators. The similarity of Lightning Terminators to natural nucleotides provides for a more accurate and faster incorporation using natural DNA polymerases. Conventional terminators often require the use of a slower, less accurate modified polymerase which also increases costs associated with their use.
The Lightning Terminators structure allows the incorporated nucleotide to remove both functional groups in a single step resulting in increased efficiency and reduced errors. Because the terminating group lies adjacent to both the fluorescent label and the custom-designed cleavage site, both groups are removed at the same time resulting in a quicker, more robust cleavage with less residual functional groups. When residual functional groups remain, the efficiency of subsequent incorporation is reduced. The presence of separate cleavage sites, such as those on conventional terminators, increases the opportunities of inefficiency. However, the single-site design of Lightning Terminators reduces the inefficiencies resulting in potentially longer read-lengths. Additionally, the Lightning Terminators cleavage site is designed to respond to photochemical cleavage thereby reducing the need for expensive enzymes.
Upon cleavage, the Lightning Terminators are reverted into nucleotides with essentially no lasting damage to the growing DNA strand in contrast to conventional terminators that leave ‘molecular scarring’. The conserved structure of the incorporated nucleotides allows for faster incorporation of subsequent nucleotides and lessens negative impact to the polymerase resulting in potentially longer read-lengths.
LaserGen exclusively owns the IP for these Lightning Terminators.