Team:Nevada/Notebook/Month1

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Contents

May 20

The whole team met to discuss project goals, human practice ideas, and training concerns. We made preliminary plans for how to divide the work and exchanged contact information so that everyone could keep in touch.

May 21

The new team members met with Dr. Howard to learn the basics of making plates and antibiotic stocks, operating the autoclave, and doing dishes.

Plates with the Erwinia endolysin, Xanthomonas endolysin, and ice nuclease target sequence were streaked out.

May 22

The team met to go over pBAD cloning and expression protocols as well as primer design strategies.

We prepared liquid cultures of the Erwinia endolysin, Xanthomonas endolysin, and ice nuclease target sequence.

May20–26

Monday The whole team met to discuss project goals, human practice ideas, and training concerns. We made preliminary plans for how to divide the work and exchanged contact information so that everyone could keep in touch.

Tuesday The new team members met with Dr. Howard to learn the basics of making plates and antibiotic stocks, operating the autoclave, and doing dishes. Plates with the Erwinia endolysin, Xanthomonas endolysin, and ice nuclease target sequence were streaked out.

Wednesday The team met to go over pBAD cloning and expression protocols as well as primer design strategies. We prepared liquid cultures of the Erwinia endolysin, Xanthomonas endolysin, and ice nuclease target sequence.

Thursday Glycerol Stocks of the cultures. (9 in total) Miniprep of the 3 liquid cultures and Nanadrop analysis of Plasmid, Liquid cultures were prepared again but with new KM antibiotic. The LB Kan plates were checked and it was found that the Kan was bad. New LB Kan liquid was used to start cultures for minipreps.

Friday Nanodrop of the 3 cultures again (a higher yield was obtained). New LB plates (5/24) were made. Minipreps were conducted on Erwinia endolysin, Xanthomonas endolysin, and ice nuclease target sequence genes in the Genescript (Kan+) plasmid. Submission plasmid pSB1C3 was transformed into Top10 E. coli cells.

Saturday LB plates (5/24) were checked for resistance. pSB1C3 plasmid was grown on LB CM100 liquid for minipreps the next day. Haley and Cody performed a miniprep of PSB1C3 S1-S6 liquid cell lines and of ICE,Erwinia and Xanthamonas. Cody and Haley performed a digest of PSB1C3 S1, Xanthamonas, ICE and Erwinia plasmid using restriction enzymes EcoR1 HIFI and Pst1 HIFI. Liquid culture of PSB1C3 and plates made to keep cell line.


Sunday pSB1C3 cultures were miniprep’ed to isolate DNA. Samples recovery and purity was determined using the Nanodrop spectrophotometer. DNA samples were stored in the 2013 plasmid submission box at -20oC.