Team:Georgia State/notebook


5/13 Group B: Made LB Broth and overnight cultures of pGAPzα B Group C: Made YPD broth; overnight cultures of pGAPzα C from plate 3; transformed E. coli cells with pGAPzα C and plated.

5/14 Group B: Made Miniprep. Group C: Made YPD plates with sorbitol and zeocin; miniprep on overnight culture.

5/15 Group B: Restriction digest Group C: Made eight hour cultures from plates made 5/13; made YPDS plates (no antibiotics); plated P. pastoris on YPDS plates; made YPD broth; made overnight culture of P. pastoris in YPD from glycerol stock; made overnight culture of E. coli and pGAPzα C from eight hour culture.

5/16 Group B: Analyzed the Gel; made low sodium LB Broth. Group C: Miniprep of overnight E. coli and pGAPzα C culture; restriction digest of miniprep samples 1-1, 2-1, 2-2 (cute with either EcoRI or Bgl II; ran gel at 80 V for 90 minutes with samples loaded in the following order: 1000 bp + ladder, 1-1 E, 1-1 B, 2-1 E, 2-1 B, 2-2 E, 2-2 B, 100bp + ladder; made low salt LB broth; made 200mL overnight culture from 5/15 overnight culture 2-2; made 200 mL 1M sorbitol; autoclaved 1L deionized water for use in transforming P. pastoris.

5/17 Group C: Miniprep of 200ML overnight culture from 5/16; glycerol stocks of E. coli and pGAPzα C; stored plates with P. pastoris growth in refrigerator; plated pGAPzα C and RFP and placed in incubator.

5/20 Group A: overnight culture of pGAPzα A, B, C, and A modified. Group B: miniprep of pGAPzα B. Group C: eight hour cultures of E. coli + RFP; midiprep of E. coli + RFP; overnight culture P. pastoris; digest of pGAPzα C with EcoRI and XbaI.

5/21 Group A: Made YPD broth. Group B: Miniprep of pGAPzα B. Group C: minipreop of eight hour RFP cultures from 5/20; digest of RFP with BglII; overnight culture of P. pastoris from overnight culture 5/20; digest of pGAPzα B and C with EcoRI and XbaI; precipitate pGAPzα B and C DNA; ran gel confirming digestion of RFP.

5/22 - 20°C Freezer was left open overnight Group C: Gel shouwed DNA present in pGAPzα B and C digest; digest of linker with EcoRI and NheI failed; made competent P. pastoris, glycerol stocks in -80°C