SEQUENCING REACTIONS WITH LITHIUM FOR PULSE WIDTH CONTROL

摘要

Compositions, kits, methods and systems for single molecule nucleotide sequencing comprising producing polymerase reactions having lithium that control the median pulse width for incorporated nucleotides are disclosed. The levels of lithium are used to control pulse width while allowing other sequencing parameters to remain within a desirable range.

主权项

1. A method for performing single molecule sequencing having a pulse width within a desired range comprising:
providing the composition comprising:
a modified recombinant phi-29 type DNA polymerase enzyme having at least 80% sequence similarity to SEQ ID NO: 1, a primer, a template nucleic acid, and a plurality of labeled nucleotide analogs wherein the composition comprises Li+ at a concentration of from about 0.05 mM and about 40 mM. observing the association of the labeled nucleotide analogs with a complex of the polymerase, primer, and template nucleic acid over time to determine a sequence of the template nucleic acid, wherein the concentration of Li+ is selected such that the single molecule sequencing reaction has a median pulse width between about 10 msec and about 400 msec.