Formulations and methods for stabilizing PCR reagents
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-009/12
C12N-009/96
C12Q-001/68
출원번호
US-0654457
(2013-12-20)
등록번호
US-9725703
(2017-08-08)
국제출원번호
PCT/US2013/077290
(2013-12-20)
국제공개번호
WO2014/100755
(2014-06-26)
발명자
/ 주소
Whitney, Scott E.
De Los Rios, Stephanie
Ghirmai, Senait
Muller, Rolf
Liberal, Vasco
Wilkinson, Steven P.
출원인 / 주소
Biomatrica, Inc.
대리인 / 주소
Wilson Sonsini Goodrich & Rosati
인용정보
피인용 횟수 :
2인용 특허 :
222
초록▼
Described herein are stabilized polymerase compositions comprising a polymerase and an polymerase stabilizing agent, such as a non-detergent zwitterionic stabilizer or a cationic ester disclosed, for use in nucleic acid amplification or nucleic acid sequencing. Compositions are provided for the stab
Described herein are stabilized polymerase compositions comprising a polymerase and an polymerase stabilizing agent, such as a non-detergent zwitterionic stabilizer or a cationic ester disclosed, for use in nucleic acid amplification or nucleic acid sequencing. Compositions are provided for the stabilization of one or more polymerases in a single stabilized liquid formulation. Also disclosed are methods for making and using stabilized polymerase compositions and kits for nucleic acid amplification and sequencing comprising the stabilized polymerase compositions provided.
대표청구항▼
1. A liquid composition comprising a polymerase and a storage stabilizing agent, wherein the storage stabilizing agent comprises at least one cationic ester compound selected from the group consisting of: [i] 1-(2-ethoxy-2-oxoethyl)-3-hydroxy-1-methylpiperidium bromide,[ii] N-(2-cyanoethyl)-2-ethoxy
1. A liquid composition comprising a polymerase and a storage stabilizing agent, wherein the storage stabilizing agent comprises at least one cationic ester compound selected from the group consisting of: [i] 1-(2-ethoxy-2-oxoethyl)-3-hydroxy-1-methylpiperidium bromide,[ii] N-(2-cyanoethyl)-2-ethoxy-N,N-dimethyl-2-oxoethanaminium bromide,[iii] 2-ethoxy-N,N-diethyl-N-methyl-2-oxoethanaminium bromide,[iv] N-(2-ethoxy-2-oxoethyl)-N,N-dimethylcyclohexanaminium bromide, and[v] 2-ethoxy-N-(2-(2-hydroxyethoxy)ethyl)-N,N-dimethyl-2-oxoethanaminium bromide. 2. The composition of claim 1, wherein the composition comprises a disaccharide, a trisaccharide, or a combination of a disaccharide and a trisaccharide. 3. The composition of claim 2, wherein the disaccharide is sucrose, or the trisaccharide is melezitose or raffinose. 4. The composition of claim 1, wherein the composition comprises a polymer. 5. The composition of claim 4, wherein the polymer is polyvinyl alcohol (PVA). 6. The composition of claim 1, further comprising BSA. 7. The composition of claim 1, further comprising a buffering agent. 8. The composition of claim 7, wherein the buffering agent is selected from the group consisting of Tris, MOPS, HEPES, TAPS, Bicine, Tricine, TES, PIPES, MES, and combinations thereof. 9. The composition of claim 1, wherein the polymerase retains the same activity when stored at room temperature for at least two weeks, at least 3 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 1 year compared to the activity of the polymerase when stored at a temperature of −20° C. for the same time period. 10. The composition of claim 1, further comprising one or more additional reagents selected from the group consisting of a buffering agent, reducing agent, a non-ionic detergent, a salt, at least one dNTP, a primer, and combinations thereof. 11. The composition of claim 1, comprising about 0.05-5M alanyl-glutamine, about 0.5-10% sucrose, about 0.5-10% melezitose, about 0.1-1% PVA, and about 1-25 mg/ml 2-ethoxy-N-(2(2-hydroxyethoxy)ethyl)-N,N-dimethyl-2-oxoethanaminium bromide.
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이 특허에 인용된 특허 (222)
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