zyP-5052 Rich media for auto induction Vols are ml. Add in this prdert 200m! 00m! LL 3. a 2Y media 186 464 274 “312 7424 1M Mgso4 02~ os 3004 8 1000x metals mix 02 as 30 4 8 50*5052 4~— 10 6080160, 20xNPS 10 Fe 150 200 400 ‘ntibities az needed ‘inalvol 200.8 500 1000 | 2000} 3000 4000 8000 Sterile iter f storing zy. ‘Additional ingredients are added to ZY media, so the final volume in the aliquoted bot is 10% ofthe total Gf 18.61 G/1860mI | 6/9301 9/9420 mb typtone 300 20 |" 10 aaomma yeast extract 100 10! 5 I Autoclave ~ _ 1M Mgso4 Steril 00. Is. MIX-see additional word document SOx5052 - 1x » 0.59% alycere, 0.05% glucose, 0.2% alpha-lactose filter Yoayeeroibyweght 259. 2509 26 ¥ #20 738ml 730 mt veo ot, Valucose(as dextrose) 259289 ga a ed Valaractose 10g 1008 soy finalvel 100m aL Feome add in sequence lactose Is slow to dasove, heat helps Autoctave BOX NPS~ 1%-200m 04, 25mM S04, SomM Ni, 00MM Na, SOM K 20 80mi-800mI¢ voce satste (nnay2soa Gey ge ge (ten) GesneApaKt2PO8 1389 1369 NaghOs Conydious) 1829 Nag setiumprsyit WS pina vol 400 mt = adi in sequence, pH of 1X should be ~6.75 Autoctave‘Trace Metals concentration of 01x trace metals 10 support maximal growth in P- 3G. Growth in ZYP medium is nt limited by lack of tone peat Frets ce mas inte isa aempt to saturate alos any met cotsning target rote 1000s te mstals mixture (100 in 50 mM HCI) ‘Ako 36 ml sterile wat sowed in 0.14 MHCI= 00d dof cone HCl) MW Income tampa Use: of stack sli (Goa) gie lat LM FeCls6thO __ 27030 50 Mre __Tists/S00m) “ 7 1 (099 20-M.Ca 1110 106 chai 12191 10-MMn 990s)! a MZa80¢7.07 20036 10-Mzn 143 so ‘ 22 Mcaciealio 211952 -M Go 28g! [aM MGuch2t0- maa Mew 0g 0 at ‘ ‘au 7 onl ‘Autoclave the stock sfutions ofthe individual metals, except 0.1 M FeCl in 1/100 volume cone HCl ‘A brit precipitate appeared upon ation of NaySeO,, which redisolved rapidly