Patent ID: 6280977
Filing Date: 2001-08-28
Classification: C12N,C12Q

Abstract:
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; andP4 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.