Method for generating transcriptionally active DNA fragments
AssigneeGene Therapy Systems, Inc.
InventorsXiaowu Liang, Philip L. Felgner
Life of this patent
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Abstract
A method for producing transcriptionally active DNA molecules, comprising (PCR) amplification of said DNA molecule in the presence of a first DNA fragment (F1), second DNA fragment (F2), first primer (P1), a second primer (P2), a third primer (P3), and a fourth primer (P4) wherein: F1 comprises a promoter sequence; F2 comprises a terminator sequence; P1 is complementary to the 5′ end of F1; P2 is complementary to the 5′ end of F2; P3 comprises a first region complementary to the 3′ end of F1 and a second region complementary to the 5′ end of said DNA molecule; P4 comprises a first region complementary to the 3′ end of F2 and a second region complementary to the 3′ end of said DNA molecule.
Claims12
A method of generating a transcriptionally active DNA molecule, comprising polymerase chain reaction (PCR) amplification of said DNA molecule in the presence of a first DNA fragment (F1), second DNA fragment (F2), first primer (P1), a second primer (P2), a third primer (P3), and a fourth primer (P4) wherein:
- F1 comprises a promoter sequence;
- F2 comprises a terminator sequence;
- P1 is complementary to the 5′ end of F1;
- P2 is complementary to the 5′ end of F2;
- P3 comprises a first region complementary to the 3′ end of F1 and a second region complementary to the 5′ end of said DNA molecule, wherein a thymidinc base immediately precedes said region of complementarity between said third primer P3 and said first DNA fragment F1;
- and
- P4 comprises a first region complementary to the 3′ end of F2 and a second region complementary to the 3′ end of said DNA molecule, whereby a transcriptionally active DNA molecule is produced by said PCR amplification.
A method of generating a transcriptionally active DNA molecule, comprising polymerase chain reaction (PCR) amplification of said DNA molecule in the presence of a first DNA fragment (F1), second DNA fragment (F2), first primer (P1), a second primer (P2), a third primer (P3), and a fourth primer (P4) wherein:
- F1 comprises a promoter sequence;
- F2 comprises a terminator sequence;
- P1 is complementary to the 5′ end of F1;
- P2 is complementary to the 5′ end of F2;
- P3 comprises a first region complementary to the 3′ end of F1 and a second region complementary to the 5′ end of said DNA molecule;
- and
- P4 comprises a first region complementary to the 3′ end of F2 and a second region complementary to the 3′ end of said DNA molecule, wherein a thymidine base immediately precedes said region of complementarity between said fourth primer P4 and said second DNA fragment F2, whereby a transcriptionally active DNA molecule is produced by said PCR amplification.
Dependencies are inferred from the claim text.