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Teamttn Co aes Va pro A [3,6 spre, SHH This procedure is obviously not working pmmalkmbihy properly, presumably because of confusion between mtants within an inoculum to a plate, and those which occur during the growth of coloniew on the plate. The samples from homologous sites are probably too small to have a reasonable probability of carrying an identified clone to the next inoculum. The procedure should be modified as follows: A broth culture should be spread or streaked on plain agar, and permitted to grow for just a few generations. printed to The thinly grown plate should then be icyranmfmrrxr selective and plain agar, and the clones identified. The homologuss site should then be inoculated to a small volume of broth, which is then restreaked in the same manner. puller ju aan SS 2 Jiscee MG /7zo7/ A Wi77 ~ 1632 EMS Lac. PB WIb3>xK 16327 A + _— B + — 7 30 187 18 2 27 a Bo 12 33 * “foo Po dae + oT 14 2¢ woo SEtoe Lona _. . c tacr- 4e née | 162 Ficla Aah ankarn G EMStac. RB, )oo+ . 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Hall S MK TI! _ ee No + m7~” mm A 7" a - WNC AK HY ey ERY EN MDE CUWHYYEN SH Fehr eee deli teh i aeed dd get ell pyr | Se MEMCLINNAYN LYRE CXCNnnnNX~ eT TUN YH cere nnauutse teh eeeged Lil bil teeter eee lial li ii xe Vey Per PEL Ch Cte ep pee te eee eee ten ed | a | | on +. yer 8357 ( Vass tun 2-4) ' = W /6y?7 ~§39q (Uurotn /6P) = WY 164% hian [2A % YOR = ea SAD a 8x 1d = Sao. “sy seed eg uett!) purrtawy ROEY « allin ao, . e EUS he wt 1 ual /, a (ere ! x yew ' Vins vet Va ER ' Le frae, riMnctge LR -.- | , ; tw’ bof~ ‘ap cee 2?) ud (647 uv | Sy - nud m. 6 pele | ranstly Corrota/ 7 lac- get (§). Hn - Clr - yuc + Pel -| x Kgl alr ~ er A oth maeh ecto | oj pon" ee ial P Daf - SS ). I Bis. 7 Yh. /t /2 zt £. 12 yo PAY vurak Porn ee, tT Porte tha. AD tel te AL f. N. tt, po 3 S unto. funk _ , > (y) a fF. Lill. | then 2 ; 3S thn. wi7? potatos - | u 3 4 A. PIL [wk mn ENGhee ist ” | ; " t Sul WY 77 6 Plot. TR om OP Larsen 7 hore [ork Penner ty , Gylwaws : Fe ub, url onter ; 6 stave ca SD bre — IA, 2 tack wan de YOO Sr(bac +). Huck tbh . Ref bc. 1th Vavdhon ree Sema ee fe A Z had fF Kata ; (aA LO tniaerty, Mam ganchd effidhy heb PL fo: oN op Iso 4 rweke, S. sha L prrathAnriieo «EMS Lac 3-5. Pile wih stink ot mn EMS foes lee , Dlo).—" 3daqs: na ge fon LPeoblAs T- bt Auris Porn ng te Carey our. “o + ro fev . § hoor wWIihyG + B74 rink in Camaros . qa * J /632- oO os ” - 6 a Cardia | Me Aacdenss of qencte aitirdy vr prrvicblon tetinee f 12. TL- +-] fy é alae ge | ipa tn, FL Cesare Joly 61951. A Wid67 YM W136 2 Cc Witt ¢O 17 D: ez pte t D ome TER, ); , £778 Fpl 22 Th,. ants clonh 60 in ne FB, por faa a EArg 2, Po | Y 7 Tad ak ears eoph> mn EB loc b pr fuller chaticeln, 6 sav. cP wl TL - -B-~ s*® lac - Wed]. “w Ul § are T-L-(4,-) S% fap - Pab- X gf NH ~ saab - (s R putanK >>) 4/51. E. W1OY5 x corset ENS bac, nd, p14. Errs Lae 75 Yot A MH 10 °fo- | Halenl fron S53 Wiheads x W/632. gsr | EHSMK ff - dtrolee fins, Dio) C EMS Lac. PAL: eervrf eres Lar. - Moo cell bac — hen EAS ig ~ He | phe Foca, Dio) ef — ' yhishto & le prey alh Lac —, EX1S tne, [eewwk ports Lind Abt nad pracy bee - La farats be a. canafr ll pace MH, 10h. b. faumeafer SD lact elev Xo , Te, rl Sy 3549 EOS NGI /6¢@ 298 bE c. Ptamagen 25 lac - 3 | fit Ftal Te Tl Nok 3]isis Corde tenss ,, lec, Hof ot adap det be » ! (+) +1 ! ay fur bat 0G Cigdereruws,. + f t+ {yay 1+ — (4)? w, Aten? S Tl Tb RK ot: R : Q- R K~ KR '- oR tO R OR KR - 8 : R . iy - 26 R Rt +7 R 7 R . 4 7 4 _ 3 g nf 92 OR ~ BY R R -3 AR K- - — B&R “36 AR R 7 RK -_ R _ R. - R- Ace Cycepy 4 1z BL Emsb EMS LAR MTL Xyt > § 71 7¢ _ + RK 7 R +: & Ri (s- — oY R— Ro ? a R- R K ~ ~ R-7 ~ ~ RYT ; _ R wwe +MucoiD R ssales~ ~ ~ R-~- i—_——— R R ~ = R7- + R RK _ _ R- Jol (0A /03 /ot /0$ /06 {07 169 {07 yo (tf [2 3 ut. “es Ht uN? HS le? 120 (2 sa2 {a3 1a¢ (a5 jab / 27 sav 730 (3/ {32 {33 13h ES {37 133 140 se / #2 198 #4, 1¥5 /¢6 ie 1¢ ,¢7 (50 LAC MAL Emme tEms LA mitt Xyek S 7, 7¢ € + A) + Mucoid @) Re T — Rr 7“ ~~ R “ R . Lac MAL mt Yyt S 7, +| 152 _ o re q) ! 155 aR ot /63 Rie {70 - _ Race Ss +) +! VP : SeQn +1 EMS Lae LAC Ae mTl Xyl Ss 7) 7 202 - k- aoe - + —~R Ao RK - 21s” K boc - 8 +1 ns \ R 239 a 3 tay RK K s A. A Ye Tani e aS ML Wanna nYnan ov AAAS m we IN ” 4 > we. “a 2 * Ys “ Wag MM ae ar awa 4 a 5 + : | 2 Due? y a = a I se | f | | ¢ | { \ { | & J = [yf | 3) & ~ ~ A mG SRBWRO lac— ) uly |S, rq)! Poe iealey wd Posse rang. USA stl aay Kpucd . 1 aul 0) lar YB tae. Area I nb SI. 42° AWD we HP Pot & x Ti wget A JB: Renok « a 46 etenrer,. F! pmgtes, (pote) 2), GOP | weml able,). b. 10 elas; O mngker (oe Lote) , Le fran b sitive “et sec ada geoth. dparuk TO! url » py hroks Beet 1 Prt - 7972 FY é f . : o «ff & 4. Park jo? 777. Are 1 DluckK ! IS’ clonee ! 4 Ee (3 Prete S slr. 110 at q ee | ~ f Jai bak, uh eee MIS Plate A aun betula porn i ~ Sie an cecbation : ‘ dee and topes tener bibl with TI 29 sperurt rman all sores . Pele \ clne = 1 a“ B po Seni caplet | Tela} Wo semghee | | Nei, > Kener = 2-4 tnt dt Seo out : a aeotHin, ZY he, . _— aT . . 5 y “ bale — ~ a Ls. Set, “Marek, oak 1" B5 ES. duoc t tadofer 2b, to eal il & Ine 1 a pow pape F cotlemensel 7 CA Tpp oth) were reine, Wa palin . buth aus nar delta, Arve . 0 denver, At 854 fr -swprblon, fyprwrel appt. Tl wer MrhirlA Selector : W-n2A/FT}), fs 17, 165). Octet Y angle colony beth ethic Aly, ~859-I | PGR EMP ke nate a Shes Pot & TI inte Of mn ° S hore; Tiel: 16; pe Pr? N18. Pile chan bir te Pernramoay. é 3-8/1. 407 " 10°? /0 Tink glad m E11 Bore By Punt t TI pble Pvert ele» Fro, Ali. &§ 23 chyvw, 1[-3 aay | | | eh AL - >» ° ye Purnia rrey. O54 3 v * 7 . - . N14. h herr trite poctrtt, tg 3 wo?) fi Aik wm & 4D bee_ V4 —- 10°? 30°77 Jo > g °° A20. C. lca boo cells ) No v8 TT Prk Lotestian BD B: Aclones 1,0 singles | ae a re A: th clones ; 2,0 Peajles- \ “ ve i 4 | JOAR Jo’ tbh, — 3ptetice © MA Dhow, meyre cyrle’y fewn/ A Stale ub Y of He,2. y, Oe BD wtte; 193 wOP Tet “Th. y Mace uarslp J,” a vow. Cc 0 Lt . rs / . | bith , SAB... ” Go Fo—L 7. Cael fared vty /0 Sitfules Pure eee : art - i s/s/51. Chk ret a7 EMG gla Repheen te Eytlec. TI. phe role ut u,%. WES cle ade pole Total tow : 8 5P es 4. Aq 0 13) a 4.7) JDO vb 4 166 — G3 447 Pnyxb SG anid ckrcer By p45, Lo) /, 2 PN. wpa (PSF - 1) 6.02 wk + Tl arson. 4° 38 La tbea&® | ue Riga loca tA TY fret Arr bra toh afta pueutars, 2 Bx. . | ew 4 de IS Y O ) les S30 aa oO 6 . 3b iy jo Soa 2bh log qo Diihgl en senc> feme, 2b fla Mae veh aL 26 fr pullirte) fp Bree hoe “pt. Worn choral “ ,. c! {mk mapa (Emr) ata tol cL Imd Cake terete {ml » 10:1) Clones u LZL i) VW ws vv) Vv —— —yv_ vy wry Vv Vv VU ¥, is v % ye ver v ve v Ww wd WAY v “view YY w--—-wv v® @@ -~~-vG@D - - v yw MY ow wy oe vu VVy = v Vv Vv uyY™ wy Ww wy Gay vue 56 2 OGY 244 VWVvV -w v SO VY WW vy wv y v voy U 3) viv —wiiv \ aA WY wiv SBP>vvy Vv ws av 4Y v B35 w Vv v- 3S6~vV Vv 37 WY wwii BF weeow a, UV 34v ov Yo WY ~. Woe - Hf Wu 92 purr G2 ue Y¥ Vw w YY Ww Ub Wy MY WY wo ® Y 7 wer pur YS we Yun vu wwe UP wy Wy m—mowy SUV vyvu v v wiv Ss) wo Wm o> vw yy Ww oe Y YY — 53a Ow © w a oS VV we ogee SEQ SS Vv Vv Vv v “ STAY Y WX ow SF UVOv 54 @v — twerer IS, 1,29,10 IT, 16 S29 792), 1A Chl NGht Avi t 29, 97. gee 867 ___ §6 3 Ble, - pe 30, 19>) A Mbor-h- ts. Ak pt CU pits » ETS Lae OM h2- aug p l ptake . per x. a Lh Ax W-l Ki gGT-&- 2. fox wW-f = pat ~ nel eran - bpst— 3. fi x HY) —_ 2 - 4. fp «x HY nl : of 5 AK N40 pot bint ang. 6A xX W606 [er bh ~) rls. th ptoker Barton . Other repraes wrth W606 HWS, HI4G0 un pth, fF. AA, , Nae Sey s por Metter. > Ab free. Vrafrrzdy 2 W837 SbY 1fxe PLL. p, ENG Lae CoP e I eating hy acai ll Xe Veo baz jon rlok Lenten mm tx 20 pa » far tetris ] ca P00 mane teats fa ance, A delle cant /0x 3s0 No- auydryrhs help ! = B53 . Toth cn. 400 feoks ! Poke on EBL se. LE. Wrrhumth rveords slags 647 TF. Vacahte tape, fanvrt Mele A laer. ! z 3 ajre[st Mam scatlely € Cab ae i 1606 x H243 | wolere » AF oe 9? x sR SES Heath, lac. hi phees to Enslac, EMSlac su, fr 6° Co BDO cofeares —_ ror cag een A, ty EMSs Lac, at Siuy,. 3 WI67S X Hao. c. Wib7s * HI44 mEMS Iae | t te FL bec te N-bev aby S° els|s pond be BA lac rte s? (pute, wicot) pels »2, (72 1, a H2490x H293 M-lacvMk- TAL- 57 pey Neb S% lu * Ru b. EMIS Uk Sb So 4. Zo preed /} Laeu G tho lace Gerd) bo ” PS al~ zs 3 I 7aby a ? 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Phat 4 a ( L: ?) four 12. br Wftishy E bor fo. lage rcener a S7O largest 29 y) (997 a) Dianrliopt for PPA ( Hh -) arid PID Laue. 3 x) | UV YOsec. , erperclia! Cl ge ba ea ley hnay at howd oak lat ete I. 180 wW, /): c) t «tht ture. + (0 wf nk - cg. ho Merete gtd. WAS nt HB tre afly 10 12h, Mey ea t& NOL). PE Y ea IH. cme agrtec . PF IR b & = rw) * * CS VG IK? Yes z ° «<4 9 . Oo < “ads rdegdnddaod Joaadd woo hnwy ododorat-ddds Qadddoeda Tame las nw EL ELQ Haase re egeda TIP ach ag Q FSPSsSSES|9 Dory) Sie op c S ® (yy Q S & PRs wih Ad- Recbctes ba, cuphecn hue Teopt,~ Dercark Theee , parka HA— (pur alt wll ee oz ft ) apr PFE. / () im , Joop Mor f f raves aad pore au hy Fr tory . rp tint #1 t go : PF/2: 179 ME 1+: uv yp %» AB A> 21 HCV 22 Ay AY PL F: 23 Ad Nt Az woogd Ad A> ve \ ae ae Ay AY 6 ae Al 2) iA Al 18 YWA. N+ YA. 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"” " Jee lH fifa Ss Hive bn 340, | _ 4 os Pear lore ~ pad Menton i Pi bres. 7. RB! 12) u 187 19 x10? 2.2. 9d) } f efisfst Bl ge 20 o | tel ot / piu >, /?) yf. i ‘ é ~ . Mevef | Leryn Pou Baan | gete. r, uy C4 Aah yy Dey, 2- tay 4. ce. G0 SE = dot Mee Simkar seriyele + ) pte li erin de + ( Me , tho, . I/y0 , af Jotz )? eer 7 ‘ / cf ‘7 f of — . w a ae pelican frAree [Net a of DUM ny rR TE 2, eae a y us ~ halal p SP 164 OF 34x 105 Natl jo Jy [5 4 . ; L5y sot wen) w-77 148 forse used wm Qoorroe, 09 O08 -09~UG. Wess kdb ceerul) 1o/u fs ( Lacle ); YO -08-09.22€ yo/ Mas [OD y134¥ 10 7/2. PB. eR. Haunt jo:/, OF wl | fete. Artery mb 3.5410" puhh B. Ww Vlg Pte LUA fede Zs [Onl Pele jo™ Fo Yin kaw Kner Sarah aad 2e sad | Department of Genetics University of Wisconsin Madison 6, Wisconsin January 1, 1952 Preservation of Bacterial Cultures on Silica Gel This circular was written in response to a number of inquiries. Judging from these, present methods for preservation of bacterial cultures are not entirely satisfactory, and it would be a real contribution to laboratory tech- nique to work out a better one. Unfortunately, I can only suggest a principle that seems very plausible, but that has not yet been empirically justified. The working principles are (1) that suitably dried bacteria should survive just as well in a sealed tube under air as in vacuo, and (2) that if this is correct, chemically inert desiccants such as anhydrous silica gel could greatly facilitate the practice. The following arrangement has been tried: small vials or tubes are filled nearly full with silica gel (Davison Company, Baltimore; Grade 40, 6-16 mesh). About 1 to 1.5 gms. of silica fits well into the tubes used. The tubes are plugged, then baked in a sterilizing oven at 160-180 C., 2 hours, to dry and sterilize the tubes, These are stored in a desiccator. The bacteria to be preserved are suspended in 2% peptone. About .05 ml. is pipetted directly to the silica, and the end of the tube sealed off, The tubes were then stored in a refrigerator. The water disappears very quickly. To regenerate the cultures, the tubes were broken, and the silica poured into broth. Considerable gas is liberated. After about an hour to allow redispersion, viable counts were made on the broth, The 24-hour survival, in apparently dried condition, was quite high (about 1¢%), but this was more encouraging than long-term experiments. After four - five months, the survival has been low, of the order of 10-5, and some tubes are inviable. In its present form, the method igs not a success, and can- not be recommended for long-term preservation and storage. I think that it could be greatly improved, without complication, by experiments leading to a better suspending fluid, and possibly by drying the cells on a layer of glass bead over the silica. What is most needed is an improved theoretical understanding of the biology of suspended animation in successfully dried cultures. Despite its shortcomings, the silica gel tubes provide an ideal method for mailing cultures, They are not affected by undue cold in the way agar slants are, and probably ought to be more resistant to high temperatures as well. The mechanical strength of small sealed tubes allows them to be sent with simple padding in an ordinary envelope, and the absence of any liquid minimizes pos-— sible hazards from breakage and leakage. T hope that other workers with suitable facilities to study preservation problems, or with a potential interest in the biophysics of suspended animation may be able to make some use of this suggestion. Joshua Lederberg J PIS- eleven 25/1591. A. K-12 orennghh, ecuth. Pelle 110 (ot. 2x 10% Jon ) A2 O2u8 fesay oiginel 12. | 5, 7x70 7 AS 00> mL B2 Ii jroo (est 2¥ 708 |b). O%unk B2. OG lym. 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