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US4663283A
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US
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Method of altering double-stranded DNA
Assignee: Genentech Inc
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Filing: 1983-10-20
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Granted: 1987-05-05
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Expires: 2003-10-20
Abstract
Abstract A method of altering double stranded DNA comprising converting double stranded DNA to single stranded DNA in a region surrounding a selected cleavage site, hybridizing a complimentary primer to the single stranded region formed in the previous step, and restoring enzymatically the second strand which had been eliminated.
Inventors
- 👤 Dennis G. Kleid
- 👤 Daniel G. Yansura
- 👤 Herbert L. Heyneker
- 👤 Giuseppe F. Miozzari
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 19:40
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/71; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/61; C07K14/655; C12N15/10; C12N15/66; C12P; C12P21/00; C12P21/02; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
117 words
A method of cleaving double stranded DNA at any given point which comprises: (a) converting the double stranded DNA to single-stranded DNA in a region surrounding said point; (b) hybridizing to the single-stranded region formed in step (a) a complementary primer length of single-stranded DNA, the 5' end of the primer lying opposite the nucleotide that is 5' prior to the intended cleavage site; (c) restoring that portion of the second strand eliminated in step (a) which lies in the 3' direction from said primer by reaction with DNA polymerase in the presence of adenine, thymine, guanine and cytosine-containing deoxynucleotide triphosphates; and (d) digesting the remaining single-stranded length of DNA which protrudes beyond the intended cleavage point.
Claim 2
Independent Claim
117 words
A method of cleaving double stranded DNA at any given point which comprises: (a) converting the double stranded DNA to single-stranded DNA in a region surrounding said point; (b) hybridizing to the single-stranded region formed in step (a) a complementary primer length of single-stranded DNA, the 5' end of the primer lying opposite the nucleotide that is 5' prior to the intended cleavage site; (c) restoring that portion of the second strand eliminated in step (a) which lies in the 3' direction from said primer by reaction with DNA polymerase in the presence of adenine, thymine, guanine and cytosine-containing deoxynucleotide triphosphates; and (d) digesting the remaining single-stranded length of DNA which protrudes beyond the intended cleavage point.
Claim 3
Dependent Claim
39 words
The method of claim 1 wherein steps (c) and (d) are performed simultaneously by reaction with DNA polymerase which polymerizes in the direction of 5'→3', is exonucleolytic in the direction of 3'→5' but non-exonucleolytic in the direction of 5'→3'.
Claim 4
Dependent Claim
12 words
The method of claim 2 wherein the polymeric is Klenow Polymerase I.
References Cited (Backward Citations)
US4351901A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-05-04 | 1990-11-03 | 1991-05-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-05-04 | 1994-11-03 | 1995-05-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-05-04 | 1998-11-03 | 1999-05-05 | $7700.00 | Window Closed / Lapsed |