Paris/August 13


yesterday -- tomorrow


Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 Size Description Product Left []ng/µL Date
D41 araC/pBad-J61002 plasmid BV L1.2 (araC/pBad-J61002 plasmid) Spe1 Pst1 Vector with the pBAD promoter ready for insertion of a RBS/CDS insert August 13
D22 pSB1A2 L6.1 (rbs-CRE) EcoR1 Pst1 August 13
D33 RBS(B0030)-CreORF BI L6.2 (rbs-CRE) XbaI PstI RBS-Cre digestion for use as a BI August 13
D42 Lox66 FV L9 EcoRI XbaI pSB1A2-lox66 as a Forward vector August 13
D35 Lox71 BV L10 SpeI PstI pSB1A2-lox71 as a Backward vector August 13
D43 gfp-tripart FI MP8.1 EcoRI SpeI gfp-tri part (strongRBS-ORF-T) ready for use as a forward insert August 13

digestion mix:

  • 25µl plasmid
  • 1µl BSA
  • 10 µl NEB2
  • 60µl H2O
  • 2µl Enzyme1
  • 2µl Enzyme2


Number Insert Insert Volume (µL) Vector Vector Volume (µL) Comments Number of colonies
C1 MP3.1 digested with EcoRI (D0) 2 control of ligation efficiency ~1000
C2 D35 (L10 lox71 dig.SP) 2µl control of D35 digestion 5
C4 D42 (L9 lox66 EX) 2µl control of D42 digestion 5
L26 D33.2 (L6.2 RBS-Cre XP) 5 D41 (L1 J61002::pbad SP) 1 Insertion of RBS-Cre behind araC/pBad x100
L27 D27 (ftsZ PCR XP) 5 D41 (L1 J61002::pbad SP) 1 Insertion of ftsZ behind araC/pBad 6
L28 D32 (ftsAZ PCR XP) 5 D41 (L1 J61002::pbad SP) 1 Insertion of ftsAZ behind araC/pBad 8
L29 D43 (MP8.1 gfp-tripart ES) 5 D42 (L9 pSB1A2::lox66 EX) 2 Insertion of gfp-tripart in front of lox66 x100
L30(A&B) dapAp [O31+O32]0.2µM 2 D9.1 & D9.2 1 cloning dapAp promotor in J61002 x100

Ligation mix:

  • 2µl 10x buffer
  • 1µl Ligase
  • XµL plasmid DNA solution
  • YµL insert DNA solution
  • H2O qsp 20µL

Reaction in 20µl

ON @ 4°C