Today, early in the morning I streak colony from desired-putative recombinant into another petri-dish, so in the evening they could be inoculated into 5 ml LB containing appropriate antibiotic for plasmid isolation tomorrow.
I got enough putative clone especially from PPICZalpha-AO6 and AO5, except for PPICZB-GFP EcoRi and PPICZalpha-AO1. I remember the latter of two, its vector came from different source. AO5 and AO6 I got by myself. however, we cannot sure until we digest to verify desired clones.
I usually make LA+kanamycin (50 ug/ml) for recombinant E.coli containing kanamycine resistance. for example we we want to use 300 ml of LA we must put 300 ul kanamycine from stock 50 mg/ml.
I also streak e.coli recombinant for cryopreservation I got from my previous experiment as follows:
1. pOKT-kan4R
2. pOKT-GFP+S (EcoRI)
3. pOKPldhSlpA-AO1
4. pOKPldhSlpA-A05
5. pOKPldhSlpA-A06
In the evening, it’s about 3.30 pm I prepare for inoculating putative recombinant that I have streak early this morning ( we select 6 of each petri dish representing all of clones) for plasmid isolation tomorrow.
some previous clones must be refresh today:
1. pGEMT ICPPFN
2. pAS900
3. pGEMTGFPUV+S1
4. pOKT-GFPUV+S1
5. pOKT-GFP+S (EcoRI)
Zeocin (25 ug/ml) in 5 ml LB, so we have to add 2.5 ul Zeocin from stock 100 mg/ml.
