IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-3

=Tandem OriT by Qu Mingzhi=

Amplification Culture of F-OriT->pSB1A2

 * Culture at Amp+ plate for 12 hours.
 * select Positive OriT-pSB1A2 Colonies from Plate,Culture in liquid LB,waiting for mini-prep.
 * pSB1A2 self-ligation nagetive-control did not grow.

=Lock & Key By Yu Tao=

=oriT Knock Out=
 * By Xu Anting

Sequencing results

 * Got sequencing-corrected strains of F and R751.
 * All pSC101 sent for sequencing (7 colonies) have the same point mutations in several regions. I wonder if our strain from CGMCC is different from that in NCBI database.  I will go on with these strains and meanwhile repeat the oriT-pSC101-deleted construction.

Amplification culture of FRS

 * Amplification culture of sequenced strains: F (No. 3); R751 (No. 3); pSC101 (No. 4). Shake with LB-Amp+ liquid in 37℃ for 16 hours.