2) —KLG60 1236 cn pe ss eee he Sikeg Ree fm r./I 7 s . thoy GocLp HM Ge cath, 6 srgnsft alls 37 he | j ont pont ie Bate he po eg at by ote Ht I gave uv, tantly foLr Maer 4/v22), . 3 Llacen peal on nese musmtsveen wee " Theee ume J237 ee MAR 11 1955 | “8h . . — [wen . | Lb tah — : . : | a sd OT ¢(2,3..) 1, —, # shod 20 2 F238 BT | : 3 *F o (2°3' oy 2] 20 Bs 7 yr os 2 sf $49 Pid? —w Sh Moz. as forston8 Na 40 855 elf / L32 big cl pbo (dat fk < Phe i nat) A. Fesckepoit=. ddlele 6232] $2 by Wan, BD) Dawe fab) (bln frat of HL. b bale pooD te rte. 7a Auk Lule (= 0,%,10 cll, on Hoh fs fan Se 1 OSop4. — HP lament tx f° tad Le one Sass, y Be elle - 2B he oar. » eibrard: So fi '$ ce ie he . ° i MAR 11 1955 . ‘e be 7G eee /. | A, Conteh Mod ue Ba, ot Boa ff hed seman id howe? js. » haste 5 main eon len, ng il 548 ahr most may 7 he [2 T: dp oe TRAILS i, BI, | ( at * Basdhitids , at /7 Beg | OF — offo Leatuwet (03%/ ) fs - ait Sern ¢ atesdinaf) FI Nove AN Sb alse “dae sonuve 5 ules fe 20% mine / [orekt Mn voeehod Bor catinl e cope dur | J. o | [Oo 10 7 \ Tt we “ / we f - MAR 1 2 1955 i (bb pipe? Wo troLe . MAR 1 2.1955 Miiabed © d Sd+4 RK t6- Cre Usa ef ; rites Covet i 2] +6: fe “ate e antonel , 2 ZS 3b Y- a | a OFLVST- . | | | | 2% ye TH. . (27- : ae usb br flaw - ff fn mebatit peecleg - Pexbicibe ap pts ~ te se Sofeke Sy. de, aabtionr (oesgpinel MAR 13 = a - pipatie LEG cay stepuntd uly? thes. Co . ye Th 0a tf wmted. MAR 12 1955 Bearings: — Since Jan 1, I have been primarily occupied with Salmonella trails. An important ques-— tion-has been the uniqueness of the"Z particle". This would be hard to establish by quantitative data on the clones directly, and I have been principally concerned with looking at platings of small clones in motility ggar. The xwmekexkk results with SW-967 are not fully reliable owing to spontaneoys “minor trails". This work has been done onay since Febr. 23. Before that, from Jan. 11,I was mostly fiddling around. I mist have been preoccupied with otherkinds of experiments __t00, or weiting or what not, since rehatively few experiments are recorded. There are also some experiments on crosses of heterogenotes, but DCG did most of the routine on these. From Jan-Febr., there were a number of misc, experiments on conditions of : . plating ete., which amounted to very little. There were some indications of major and minor trails. Also developed technique of trapping from conc. cell susp. _ Summary of clone platings. . (trails per clones}/dfd per platings) and comments 1227: 1/20/25 T: uniqe ° —x 967 12/60/74 All major trails unkime. some djst. mass pla. 1228 3/9/10 : majors unkdwa, noticed addl minors 1%/86/ 100? 1229 8/31/39 All T unique 24,/198/200 OO . 12290: apont minors . , : _ Non-linear however. Confusion with | spontaneous minors. ee ©. 0 at ERIE caer ree. —x S666 “Almost as prolific source of mtiles ~Izgz «4 /18/25° Single mors, pot otter clusters. 2159/22 * Few, 5$w/13/79/100--- Clone size c. owe a wn 1233 1/34/47 Single small trails; a few other ——__——--— - -glusters. c. 26 Lye a ene nena 1234. 6/25/48 Definite concurrence of smaller trails or larger 30/149/200 cluste 33, hayd to define. Av.CI.S. 6. 23-4. 3 sw. 1236 57 clones, follow micrgose. plate only 3 NVG. . 18 el., 7 aw. 1237 (eff. Tween) This prepn. seepsng. 1/ 173 fies | wo ee a although very fresh. . 15 sw. — Should compare directly with 1234 prepn. __._48 clones. followed micros. 4 gave. sw (segr. non mt.) ee 10 quédéscent clones gave only singles; 6 with fairly numerous motiles gave 2 T's, + 1 with clusters. MAR 12 1955 The principal point is perhaps best met by experiments like 1234, plating fairly early. A corralation of trails with pluricatenates like 1237 might be worthwhile, but more laboraious. In view of sluggish motility of early log phase cells, this should perkkps be done with earlier clomes in aged medium. In some prelim, expts. yesterday, I noticed that TM2 transferred to aged medium supernate was more actively motile, particularly showing a more jerky motion with shorter fgee “path. Examination of TH2 in motility agar suggests that many cells are directly immob- dlized, others move in apparent interstices, but still more slowly than nobmal. There must be a considerable accidental factor, and cell with numerous mobile progény naturally has best chance to propagate a trail. Since genetically competent TM2 are immobilized, there can hardly be immediate correlation of genotype amt (or plurieatentation) and ability to move. Should watch traiza in situ if possible. Why not? Plans: continue with experiments like 1234. Compare this prepn. with 1237 in yield of trails. Continue with medium effects. Set up trails in situ, selectédg cells which remiin motile in gel. to see if these form the most chains. Do not forget many other carryovers? ~ 7 G&C - Gal, Nelson | . hetehogenote cro p) GAN Sabot, e ogenote ¢ eases | AL | J Hfr x F- DATE: vate. Whe I ( Wed )_ ner. 4 | 5 6 7 8 9 10 War Hepa: (an —H/ ea disyar Jd >= ee cll. (520) oi SHR PURE > Sal Sirigo t. 2S nf). Fgpucds rl tte Bok Zz |e 2 S323 2(¥,.) S | To 2 = Agrer>), Pe scp (¢, Om: |S HPA ) ted bef NOD py he Le grleameb Lliccharr ( lh >) gh Feodeaon MW 4) he i fb clnunealf, é es .2O YA?22/ 557 I2394: (03 G/ /Y0 144 67 117 4/ tT /§/ 37 Ns S uf 92 19¢ ST 1Z2A3BEODO. 22 3¢ 4 Oo /s S 2 13 44 12398: 97 ——— 225 134 leo lor Quite tuaks “C’ 3/ DATE: Mase 1G! Eat 7 Laet cor apt A Tod Ayal. Aaiglea. Gre et, D- NA fer) | 4. 7 dn tly, op J. ueA (jor). f om 56> ’ one i i tilayr Mc ap lel —| 1) SiHyptttats ¢ b hbtin . Z L . 2 Y , Ay TRE | — f Wnstt®D 230 Z 7 L iMCA (pes) (9 Mak rae h ewes) Z E ge . M . , , Fe, as p 42) ie for ) ¢ canagtenY etre Let ® tists tc eet po ff pe fe | et a a A (Cf w), | si ' pL, ed hee Gres ; Sava IK? Avtef soos ? SC , ci? > AA f 45%) Bsers (FT, [Center Ss Bs fib pew ‘eget (a) 7 | mons (Lag an tae ank in lin i 5) Aly 4 a hn rune, 3 oe hyve h auth lor ca. \F LL cle AM nl yon Aayal baal Moxy be Anel te pidigey ie ‘re ul retest , ASO r iy hot tl GMT chet 3LHT*) + tT che ele TE ce? S| (1 C ; rr 10), qrac}| & 6 ( (63, 74, Shae , Jee, D.. | 24 “fp 4) “ “yo. yA 2B TICE ATP | °P 12 ( wif 2) bo. [t ’ i | wn mg ops —~S* t) AN lv, ager eq. lo sage Ges bee rv / nee / )* ; Q thenle /O haw j, 5 slat hackle ~~ Ick; af Cal t- ie pe bs" pe oe 5D ener 23 wl 5 » 14 tachs feels. 8p te. Wick ? I Liman O 1) corks @ Rr, t figchy . OQ teks ] Conclusions: 193840. MAR 21 4955 among motile initials plated 1. Incidence of trails/varies with the fluidity of agar. Addition of 40% diluent gives very high incidence. In any event, agar that is hardening tends to be quite inhomogeneous, if maintained at critical tempseature. 2. Single clones can give at least one trail + large clusters in harder agar, and numerous trails in sofeér. This is clairly an unreliable criterion for singularity of catenation of higher mat order. Further plans: 1. A few more tests of fluadity and related variables for photographic documentation. 2. Shift studies to direct pedigrees; need some further data on irritants; inh. of - cell size growth and chamotactics. . 3. EM transfere. ales Tal douoll mohsne. / Bag ‘.. ve) hal eprint sdiglar- heater DS - Seams vi oad tal Mi prraming yn” fafa Ln nok flow wd, prdiyerd sro Han 3- fof rg ble ab allan, sate “belay ed Tg athens O Siam ofict ( Sra Geren milf my | fy . bg wore) Of dat: sl, < atifbesnth be, te bie. (omithn ot) APR 1 1955 Vharede Arann. [a Hf ce LMA (E a?) ~Xb66 wad : (e) Vor prgn. Vo fofeg =} fo — Jy!® fps Ee F (hlitan bh beth» Dio) / cg [LOl nh Ob Lp , x , lt 6 Vb - J2 Y/ hf. ty 7 ! Jost dy aged |-Y. | B ‘ . inary en 7 Jo 0! da eon (i (b a Cie) bah 3 furan mea! A<& @ 7? auth forbs b b 1 | ferrencouy ax SH 666 1242 30 | Penassay gives at least one not re - 0%" broth for plating clones. @14 had without further C5 had (Ine. 2 about 29+/1000 C13 growth. 10+/-00. [Plant ind 12:30 — 3300 Pa, Pl - 8) 100 Fla~, (42 3 |had 2+ 4 66 12/. ividually, pick td ate in MGA-40). (Spent broth = overnigh refr. for aml yes. (dil. MGA very soupy! | and decant. Numerous tine i still Note as many. 23/1000 t clones ) are limited to 18 clones (in =e broth czaniaeds 5 wd 1+/c. uh, of which 4 isolated to él, 2,3 9 each teol. . (Tange wicks le + broth for clones, (es- 1-4 vtag Fi sent). > While | and plated be 2/0). pate: APR 5 1999 REF: | 1 2 3 4 5 6 7 8 9 10 New . a): from aerated § » b) from unaerated culture. Both #1: 1 aiial ps2 11235-12210 Refr. | (Incub. in rot.) Note: Rotator fo ! now standard for -asration. | A): Prepn. a). Fuse traps 12:30, collect ca. 100 motilas, but use to spat (2) . 2~2215 ‘ 10) = BCG! pick to i Incub. in .5/ml Ponaspay to c. 6:15 PM, Plate in MGA # 40% KSB as diluent. B): Same collection, plant in spent broth (SW666 Aer.) to of. total clon size. St. ET. ch: Prepn. b) Fuse 4 PM. Collect 5230-6100 (some needle tr.) This prepn| as least as | good as a, probably better. 1000 (+) / 1 ml. Plate 0.1 ml samples “ire Plates; 205 in small. Compare 20 a MoA, +4 40% HSbdil., + 60% NSA dil. . 37 9 7 n RT to ey the sediment APR 6 1955 (1,7, 13, 14, 2% B I 7 7, 43 ) 3,54 >7)23,3 4,97) 307 6 e's 1S -¢. bo 1's | | AS 7 . i's 8 t!s (3-6) iT p7? (| Y oe 27 Image fed 4 qe's (a7) + 6 /-2's. poh Y, | 20-39 2 ae fucks Yaad 5. | smuhel ketal z | 4 . MAN jo 3/50 Ad's art ame 14,24, 30,14) 3, I), 192, CIV) Sure bv hr , (ai® M4 116 Aud lice | we 50 m j « pouhei). ) 37-1 5 S ¢|(<1a) 2) T's. 16 al BE o's : 18 aamesomerh Mallen, \2T 5 luda (3-1); 281 /2Y2LA DATE: REF: 10 ~X 666 )7¥2 37 7 _ APR 5 1955 () Bor pp A fem acalef iid) iy ne nauld « Aa (Sime) we Pe lhe yn of a a than Pll) — O. Fase dimge & * 7 APR? = GAM ‘Seme gous Cc, ¢/ 2 A Jo I's ; po chia, 7. | fey ole Boe fe ?, &, GoOI'S — , o34 dic HUOvs. I. ¢ fy Mir Sane | . C 3 a " ~~ dD FY CR ¢.30 15 le C - O } C- 2. Yols we T. Nehuwr 0K. : 2 DATE: APR v 1955 , REF: 1 2 3 4 5 6 7 8 9 = a x. G) +. t eS o\ eS ac Q ee Glanaan cleertic y #), an Ja, Oa 00. | gp ob DD | | 50) oD paTe: APR 6 = 1955 REF: (ap RD Tal, Mar, ok Sane par m , Cc. is (Sud S Bg |G TG SE Sy 9TH, F ee 129 O° fF yz 199 40 50 H - c+]{ D> wars rutetated . LY/YA 1955 APR 8 REF: DATE: 10 Tus Add ef roan he tY/ . _ 1, om, ! rm 5 > > ‘ ! » \ : 7 Pr ay S ; \§ a) og Qh om] © pore trredce robo USSR QRS IFO R20 Soe % ~ ian Shy 23 | Sts | | ff Tze yd — | 4 a r~< 1. 3 Ths 5 “sa . afl See A-09 1 nS: | 3 Fo Ya=-4) QO YoOSe wood Ff jo 00Q90~ > ss | IN 4 mm) : | ; ° ap& | & a PN g ® 1 > | Y Ss L | x ~ v | | | < < an | 3 3 » | : Q 3 > ~ a PrwWswr Seo tach& > x n 3 i rr APR 8 wore a S) W / 1 2 3 4 5 nod Ye = 1 3 10 VOWS 19 athe 4x phn fred fect [0+ indecle be iy opt ni) ee ape eli Bt be, bean ot 50 MO re m feb. | = 3 - é EN4/Dle ¢ ote Xe, 20 f pyre -© ~ 70 Lhe. : 1955 REF: Ay, a ft sn Je 5 7 8 9 10 We q rn * 2. = ] hae tw ATG Alyce an foerdes ‘g iy tT K ? x ba farf boc nthe ye 2 ee ly Ne tre 273 S, 4B OLR fe Iho cee oa \bAG NS | ft E17 -& UF Cau TN Ahive Ae 4 20|| ™ 4,4 ctu! By punto | We, THY bog || OM Ao” eth + 6. rr #3 é ara 30 /v 3h 2, / 4 | yr¥2C - Oo - oO, 2s _ be J) i Ya r a fun 7 Ne 50 ; . aT a8! sa! ¥Adleyup hd . - ¥ OT te 38t a oe 15, 7, iT fo! aT . * BT sc so0's BOD 7T 10 3BN's ee BE IEE MT 334. att AIT $C 371% ( Cow X tanger z) pl gels, Bot F1* 327 and ee ee ee AAS? A ¢ YY 31 2. a Sh . APR 9 1955 pyre oT ne i, (ele an | Moet Moa -2 o kelp wT. Yo,be ieee TS, meaery yom ae ; ne [vf>. bapa pg, pets nuded» Stace fterron, toy Lbs: (/- 6) jul (ebb be | foncsee? prettier. ith tile ptt ae MEA fo- So 7 We Ste clones OTS ye ry, “pe or / ee Jaw - oR c y/h0 S)>7 (ee Wan, elle, “tg ag ae glen e woke Poo ases Fact pet Av ees E ptrse Itawt eon LLY b Hanns mt tant of Paty )? | 7 aa / / a | Wh vig avr bea a \ ef k+s] uy | elana