Lab book of the gilestro lab
Batch makes about 40 plates. Making the LB Agar Add 250 mL of dH2O to a graduated cylinder. Weigh out 20g of premix LB Agar powder (VWR DF0445-17) or: 5.0 g tryptone 2.5 g yeast extract 5.0 g NaCl 7.5 g agar Mix powder well to bring into solution Add dH2O to total volume of … Continue reading LB Agar Plates
Topo TA cloning is done following loose suggestions of the manufacturer. We currently adopt the kit K204040 from Invitrogen, which is not based on topoisomerase ligation but regular T4 ligation. Comes with “One Shot® aTOP10 Chemically Competent E. coli”. Briefly. Quantify PCR product. The kit suggests to use 10ng of DNA. We usually skip this … Continue reading TOPO TA Cloning