公开(公告)号：US20150050660A1

公开(公告)日：2015-02-19

申请号：US14466577

申请日：2014-08-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Ayoub RASHTCHIAN ,  Joseph Solus

IPC: C12N9/12 ,  C12Q1/68 ,  C12P19/34

CPC classification number: C12N9/1252 ,  C12N9/96 ,  C12P19/34 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2527/149 ,  C12Q2527/137 ,  C12Q2527/125

Abstract: The present invention is directed to compositions comprising mixtures of reagents, including thermostable enzymes (e.g., thermostable DNA polymerases), buffers, cofactors and other components, suitable for immediate use in nucleic acid amplification or sequencing techniques without dilution or addition of further components. The compositions contain no stablizing agents (e.g., glycerol or serum albumin) and unexpectedly maintain activity for extended periods of time upon storage at temperatures above freezing. These compositions are useful, alone or in the form of kits, for nucleic acid amplification (e.g., by the Polymerase Chain Reaction) and sequencing (e.g., by dideoxy or “Sanger” sequencing), or for any procedure utilizing thermostable DNA polymerases in a variety of medical, forensic and agricultural applications. In particular, the compositions and methods are useful for amplifying and sequencing nucleic acid molecules that are larger than about 7 kilobases in size.

Abstract translation: 本发明涉及包含试剂混合物的组合物，包括热稳定酶（例如热稳定的DNA聚合酶），缓冲液，辅因子和其它组分，适于立即用于核酸扩增或测序技术而不稀释或添加其它组分。 组合物不含稳定剂（例如甘油或血清白蛋白），并且在高于冰点的温度下储存时，意外地保持活性延长的时间。 这些组合物单独或以试剂盒的形式用于核酸扩增（例如，通过聚合酶链式反应）和测序（例如，通过双脱氧或“Sanger”测序），或者对于使用热稳定性DNA聚合酶 各种医疗，法医和农业应用。 特别地，组合物和方法可用于扩增和测序尺寸大于约7千碱基的核酸分子。

公开(公告)号：US20150017688A1

公开(公告)日：2015-01-15

申请号：US14498827

申请日：2014-09-26

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Jun LEE ,  Ayoub RASHTCHIAN

IPC: C12P19/34 ,  C12N9/12

CPC classification number: C12P19/34 ,  C12N9/1276 ,  C12N15/1096 ,  C12Q1/6846 ,  C12Q2527/149 ,  C12Q2527/125 ,  C12Q2521/107

Abstract: The present invention is directed to compositions and methods useful for the amplification of nucleic acid molecules by reverse transcriptase-polymerase chain reaction (RT-PCR). Specifically, the invention provides compositions and methods for the amplification of nucleic acid molecules in a simplified one- or two-step RT-PCR procedure using combinations of reverse transcriptase and thermostable DNA polymerase enzymes in conjunction with sulfur-containing molecules or acetate-containing molecules (or combinations of such sulfur-containing molecules and acetate-containing molecules), and optionally bovine serum albumin. The invention thus facilitates the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules. The invention also is useful in the rapid production and amplification of cDNAs which may be used for a variety of industrial, medical and forensic purposes.

Abstract translation: 本发明涉及通过逆转录酶 - 聚合酶链反应（RT-PCR）扩增核酸分子的组合物和方法。 具体地，本发明提供了使用逆转录酶和热稳定DNA聚合酶与含硫分子或含乙酸酯分子结合的简化的一步或两步RT-PCR方法扩增核酸分子的组合物和方法 （或这种含硫分子和含乙酸酯的分子的组合）和任选的牛血清白蛋白。 因此，本发明促进了核酸分子的快速和有效的扩增以及RNA分子的检测和定量。 本发明也可用于快速生产和扩增可用于各种工业，医学和法医目的的cDNA。