Next Generation Sequencing

Quick reaction kinetics and conserved structures for accurate sequencing

Lightning Terminators™ were specifically designed to provide more efficient and more accurate DNA sequencing.  Their effectiveness has been proven for both next-generation sequencing and single-molecule sequencing using our in-house developed prototype instruments. Lasergen’s sequencing technology is based on cyclic reversible termination (CRT) which is a proven method that shows the most accurate NGS results; however, current CRT platforms do not have the advantage of Lightning Terminators chemistry. 

Efficient Incorporation and Termination

Lightning Terminators have better incorporation properties due to a preserved natural nucleoside structure.  The primary advantage of Lightning Terminators over conventional terminators is the unblocked ribose 3’-OH which allows the polymerase enzyme to easily incorporate the nucleotide in the growing DNA strand without customized enzymes.  Therefore, Lightning Terminators can be used for accurate sequencing with commercially available enzymes.

The terminating group on Lightning Terminators was designed to provide complete termination while retaining the unblocked 3’-OH.  Lasergen chemists engineered a terminating structure that does not rely on the presence of the fluorescent dye.  Therefore, Lighting Terminators have the ability to work in a number of configurations, unlike other 3’-OH unblocked terminators that rely on a bulky linker and dye label to initiate termination.  This gives the Lightning Terminators the advantage of accurate sequencing that is not dependent on the dye label.

Quick and Robust Cleavage

Because the terminating group lies between the fluorescent label and the custom-designed cleavage site, both groups are removed in a single step resulting in a quicker, more robust cleavage with less residual functional groups. In contrast, conventional terminators have separate cleavage sites which increases the opportunities of inefficiency.  The single-site design of Lightning Terminators reduces inefficiencies resulting in increased accuracy.

The Lightning Terminators respond to a quick burst of UV-light to initiate cleavage of the terminating group.  The optimized cleavage protocol provides efficient removal of the terminating group at conditions that do not damage the DNA template. This unique cleavage mechanism eliminates the need for harsh chemicals that strip the sequencing enzyme resulting in cost savings due to reduced cycle times and lower enzyme requirements.