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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.