Skip to main content

Advertisement

Table 1 PCR reactions used to map intron insertions in DFCT3, DFCT9, DFCT13, and DFCT16

From: Use of aminoglycoside 3′ adenyltransferase as a selection marker for Chlamydia trachomatis intron-mutagenesis and in vivo intron stability

Reaction Primers Target Notes
1 V1F–V1R rsbV1  Confirms intron insertion (primers flank insertion site)
2 incAF–incAR incA Confirms intron insertion (primers flank insertion site)
3 hyp08F–hyp08R cryptic plasmid  Confirms that transformation has not resulted in loss of the native, cryptic plasmid
4 incAF–GIIR Connects the 5′ region of incA to the intron Confirms intron insertion and orientation
5 incAR–GIIF Connects the 3′ region of incA to the intron Confirms intron insertion and orientation
6 GIIF–GIIR Intron Confirms intron presence
7 incAF–aadAR Connects incA to the aadA marker Confirms marker type
8 catF–catR Intron donor plasmid Confirms loss of intron donor plasmid
9 V1F–GIIF Connects the 5′ region of rsbV1 to the intron Confirms intron insertion and orientation
10 V1R2–GIIR Connects the 3′ region of rsbV1 to the intron Confirms intron insertion and orientation
11 blaR2–VIR Connects rsbV1 to the bla marker Confirms marker type
12 blaF1–blaR1 Marker Confirms presence of bla marker
13 aadAF–aadAR Marker Confirms presence of aadA marker