Ningyue Zhao

US Patent:

20020119465, Aug 29, 2002

Filed:

Aug 16, 2001

Appl. No.:

09/932256

Inventors:

Ningyue Zhao - Milpitas CA, US
Helmut Wurst - Cupertino CA, US

International Classification:

C12Q001/68
G01N033/53
G01N033/542
C12P019/34
C12N009/22

US Classification:

435/006000, 435/007900, 435/091200, 435/199000

Abstract:

Enzyme compositions, kits comprising the same and methods for their use in one-step RT-PCR are provided. The subject enzyme compositions at least include a mutant thermostable DNA polymerase and a mutant reverse transcriptase. In preferred embodiments, the mutant thermostable DNA polymerase is an N-terminal deletion mutant of Taq polymerase and the mutant reverse transcriptase is a point mutation mutant of MMLV-RT. The subject kits, in addition to the above described mutant thermostable DNA polymerase and mutant reverse transcriptase, at least include one of, and usually both of, dNTPs and a buffer composition, where the subject kits may further include additional reagents, including nucleic acids, a thermostabilizing agent, a glycine based osmolyte and the like. In practicing the subject methods, a reaction mix that at least includes template RNA, the above described mutant polymerase and reverse transcriptase, dNTPs, buffer, and nucleic acid primers is prepared. The resultant reaction mixture is maintained at a first set of reverse transcription conditions and then a second set of PCR conditions, whereby amplified amounts of DNA from a template RNA(s) are produced.

US Patent:

20020173016, Nov 21, 2002

Filed:

Mar 27, 2001

Appl. No.:

09/820085

Inventors:

Helmut Wurst - Cupertino CA, US
Ningyue Zhao - Milpitas CA, US

International Classification:

C12N011/00
C12N011/16
C12N009/22

US Classification:

435/174000, 435/199000

Abstract:

High throughput lyophilized polymerase devices and methods for their use in the production of nucleic acids using template dependent polymerase reactions are provided. The subject devices are typically made up of a multi-well substrate that includes in a least one well a lyophilized nucleic acid polymerase composition. The subject nucleic acid polymerase compositions include at least one polymerase and a carbohydrate stabilizing composition that is made up of at least one low molecular weight sugar and a starch. In many embodiments, the compositions also include buffer components and nucleotides, as well as a temperature dependent polymerase inhibitor, e.g., a polymerase specific antibody. Also provided are kits that include the subject devices.

US Patent:

63000733, Oct 9, 2001

Filed:

Oct 1, 1999

Appl. No.:

9/411351

Inventors:

Ningyue Zhao - Milpitas CA
Helmut Wurst - Cupertino CA

Assignee:

Clontech Laboratories, Inc. - Palo Alto CA

International Classification:

C12Q 168
C12N 900

US Classification:

435 6

Abstract:

Enzyme compositions, kits comprising the same and methods for their use in one-step RT-PCR are provided. The subject enzyme compositions at least include a mutant thermostable DNA polymerase and a mutant reverse transcriptase. In preferred embodiments, the mutant thermostable DNA polymerase is an N-terminal deletion mutant of Taq polymerase and the mutant reverse transcriptase is a point mutation mutant of MMLV-RT. The subject kits, in addition to the above described mutant thermostable DNA polymerase and mutant reverse transcriptase, at least include one of, and usually both of, dNTPs and a buffer composition, where the subject kits may further include additional reagents, including nucleic acids, a thermostabilizing agent, a glycine based osmolyte and the like. In practicing the subject methods, a reaction mix that at least includes template RNA, the above described mutant polymerase and reverse transcriptase, dNTPs, buffer, and nucleic acid primers is prepared. The resultant reaction mixture is maintained at a first set of reverse transcription conditions and then a second set of PCR conditions, whereby amplified amounts of DNA from a template RNA(s) are produced.