US5021335A
Expired / Lapsed
US
⏱️ 0.0 years remaining
In situ transcription in cells and tissues
Assignee: Leland Stanford Junior University
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Filing: 1988-06-17
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Granted: 1991-06-04
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Expires: 2008-06-17
Abstract
Abstract In situ transcription is provided by hybridizing cells or tissue with a primer, and extending the primer bound to any template mRNA or DNA with reverse transcriptase or DNA polymerase in the presence of labeled nucleotides, which allows for detection of cells containing the template. The resulting cDNA may be eluted and used in a polymerase chain reaction for isolation of the DNA, and/or the cDNA electrophoresed, which may provide for information concerning the sequence, or the like.
Inventors
- 👤 Laurence Tecott
- 👤 Jack D. Barchas
- 👤 James Eberwine
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 04:12
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6841; C12Q1/48
Claims Analysis
Total Claims: 3
Claim 1
Independent Claim
114 words
A method for determining the occurrence of transcription of a target DNA sequence in cells in a fixed tissue section, said method comprising: contacting said tissue section with a primer complementary to a sequence in the 3' proximal region MRNA of said DNA sequence; contacting said tissue section containing said primer with reverse transcriptase and deoxynucleotide triphosphates, wherein at least one of the triphosphates is labeled with a label to provide a detectable signal, whereby cDNA is produced by extension of said primer; washing said tissue to remove substantially all of said triphosphates present in said tissue; and detecting the presence of said cDNA in cells in said tissue by means of said label.
Claim 2
Independent Claim
114 words
A method for determining the occurrence of transcription of a target DNA sequence in cells in a fixed tissue section, said method comprising: contacting said tissue section with a primer complementary to a sequence in the 3' proximal region MRNA of said DNA sequence; contacting said tissue section containing said primer with reverse transcriptase and deoxynucleotide triphosphates, wherein at least one of the triphosphates is labeled with a label to provide a detectable signal, whereby cDNA is produced by extension of said primer; washing said tissue to remove substantially all of said triphosphates present in said tissue; and detecting the presence of said cDNA in cells in said tissue by means of said label.
Claim 3
Dependent Claim
13 words
A method according to claim 1, wherein said label is a radioactive label.
References Cited (Backward Citations)
US4886741A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-06-03 | 1994-12-03 | 1995-06-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-06-03 | 1998-12-03 | 1999-06-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-06-03 | 2002-12-03 | 2003-06-04 | $7700.00 | Window Closed / Lapsed |