公开(公告)号：US20170044506A1

公开(公告)日：2017-02-16

申请号：US15243808

申请日：2016-08-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Lei (Larry) XI ,  Roland Nagel ,  Stephen Hendricks ,  Jennifer Berkman ,  Marian Peris ,  Yulei Wang

IPC: C12N9/12 ,  C12N15/10

CPC classification number: C12N9/1276 ,  C12N15/1096 ,  C12Q2521/107 ,  C12Y207/07049

Abstract: The present invention provides compositions and methods for a reverse transcription reaction using a reversibly inactivated reverse transcriptase enzyme. The reversibly inactivated reverse transcriptase enzyme results from a chemical modification which inactivates the reverse transcriptase enzyme. The activity of the reverse transcriptase enzyme is recovered by an incubation of the reaction mixture at elevated temperature prior to, or as part of the reverse transcription reaction. The reverse transcriptase enzyme of the present invention provides for a significant reduction in non-specific reverse transcription from template nucleic acid molecules because the formulation of the reaction mixture does not support the formation of reverse transcription products prior to activation of the reverse transcriptase.

Abstract translation: 本发明提供使用可逆失活的逆转录酶进行逆转录反应的组合物和方法。 可逆灭活的逆转录酶由化学修饰产生，其使逆转录酶失活。 通过在反转录反应之前或作为逆转录反应的一部分的升高温度下温育反应物来回收逆转录酶的活性。 本发明的逆转录酶提供了来自模板核酸分子的非特异性逆转录的显着降低，因为反应混合物的配方不支持在逆转录酶激活之前形成逆转录产物。

公开(公告)号：US09476093B2

公开(公告)日：2016-10-25

申请号：US14593858

申请日：2015-01-09

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Lei Xi ,  Paul Kenney ,  Zhaochun Ma ,  Dennis Prosen ,  Stephen Hendricks ,  Roland Nagel

IPC: C12Q1/68 ,  C12P19/34 ,  C07H21/00 ,  C12N15/10

CPC classification number: C12Q1/686 ,  C07H21/00 ,  C07H21/02 ,  C12N9/1252 ,  C12N15/10 ,  C12Q1/6869 ,  C12Y207/07007

Abstract: This disclosure relates to methods of performing activation by polyphosphorolysis (APP) reactions using at least one of the polyphosphorylating agents triphosphate, polyphosphate, imidodiphosphate, thiodiphosphate (or μ-monothiopyrophosphate), and related compounds.

公开(公告)号：US20150191775A1

公开(公告)日：2015-07-09

申请号：US14593858

申请日：2015-01-09

Applicant: Life Technologies Corporation

Inventor： Lei XI ,  Paul Kenney ,  Zhaochun Ma ,  Dennis Prosen ,  Stephen Hendricks ,  Roland Nagel

IPC: C12Q1/68

CPC classification number: C12Q1/686 ,  C07H21/00 ,  C07H21/02 ,  C12N9/1252 ,  C12N15/10 ,  C12Q1/6869 ,  C12Y207/07007

Abstract: This disclosure relates to methods of performing activation by polyphosphorolysis (APP) reactions using at least one of the polyphosphorylating agents triphosphate, polyphosphate, imidodiphosphate, thiodiphosphate (or μ-monothiopyrophosphate), and related compounds.

Abstract translation: 本公开涉及使用多磷酸盐，多磷酸盐，亚氨基二磷酸盐，硫代磷酸盐（或μ-一硫代焦磷酸盐）和相关化合物中的至少一种进行多磷酸解（APP）反应进行活化的方法。