C - Chemistry – Metallurgy – 07 – H
Patent
C - Chemistry, Metallurgy
07
H
C07H 21/04 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 2286400
An improvement in adaptor-based sequence analysis is provided that addresses the problems created by self-ligation of target polynucleotides that have complementary ends. The improvement includes preparation of target polynucleotides with dephosphorylated 5' strands and the use of adaptors having a 3' blocking group. In a preferred embodiment, adaptors are ligated to target polynucleotides by a single strand, 3' blocking groups are removed, the adjacent 5' hydroxyl of the target polynucleotide is phosphorylated, and the ligation of the adaptor is completed by treatment with a ligase.
L'invention concerne une amélioration d'analyse de séquence sur la base d'adaptateurs, s'attaquant aux problèmes créés par l'auto-ligature de polynucléotides cibles possédant des extrémités complémentaires. L'amélioration concerne la préparation de polynucléotides cibles avec des brins déphosphorylés 5', ainsi que l'utilisation d'adaptateurs possédant un groupe bloquant 3'. Dans un mode de réalisation préféré, des adaptateurs sont ligaturés à des polynucléotides cibles par un brin unique, les groupes bloquants 3' sont retirés, l'hydroxy 5' contigu au polynucléotide cible est phosphorylé, et la ligature de l'adaptateur est complétée par un traitement avec une ligase.
Albrecht Glenn
Brenner Sydney
Dubridge Robert B.
Gryaznov Sergei M.
Mccurdy Sarah N.
Gowling Lafleur Henderson Llp
Lynx Therapeutics Inc.
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