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Page 1: How to grow a Leishmania culture - Jena Bioscience is l ith Hemin th ompleted m is possible ining LEXSY ultures in 10 lasks. Don´t t necessary are obtaine ilution of c igher dilutio

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All growth mLeishmania. completion wvitality the cexceeded, itweeks

For maintasuspension cculture (TC) ffaster. It is no

Best results successive doccasional hgrowth. It is incubate TCcultivate Leiscultivation foof the LEXSY

Always contrcultures are omotile. ThesMid-growth pof different scontaining sshape) and limitations afungal or oth

Keep patientwith regular They need th

If you - despsediment celincubation intransfection

Don’t centrifuby rigorous vat 2000-300pellets easier

If you cultivaaerate the cu109 cells/ml.

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e needs aeculture with ret 26°C. Higill not survive

media should Hemin is l

with Hemin thcompleted mt is possible

aining LEXSYcultures in 10flasks. Don´t ot necessary

are obtainedilution of chigher dilutio

convenient t flask uprigshmania mucor transfection

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rol appearanof drop-like sse cells arephase cultureshape. Don’tsuch subpopremain motnd must not

her contamina

t, esp. if youdoubling tim

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pite followingls 3 min at 2n ventilated T

uge Leishmavortexing. Th00g is sufficier. If required,

ate LEXSY strulture in a 1. If you intend

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erobic condegular dilutioher temperat

e at 37°C

be supplemeight-sensitivehe medium medium shoulto re-add H

Y strains for 0 ml LEXSY

use agitatedor growth-pr

ed with inoccultures of lons of stationato dilute 10 ht, loweringch longer than see chapte

nce and motshape, approe most effices always cont hesitate to

pulations. Cetile. Enhance

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rains in bior0 L fermentad to use a sti

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hmania cu

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and analysm (Cat. No.r strain mainadd serum to

ring early stnsities as the.g. 1:100 w1:50 on Moor longer inays in the saltivation for p

ells by microin size with

ansfection pulations of nlate or presecultures get

may result frof mid growby microscop

with bacteria.ension cultureonditions

counter growcarefully in fwas very hel

ed >10.000ghese procedud makes genime rather tha

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Jena • Germany • e-MFax: +49-3641-62851

ulture

n be maintaEXSY manualtes and vital

No ML-108)be cultivatedk at 4°C. Foeks. Howeved to use this

sis it is conv. ML-411, 41tenance sinc

o the BHI med

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will not adversnday and 1tervals betw

ame medium protein expre

oscopy. Cellsone flagelluand platin

non- or less merve a culturt longer androm nutrient wth culture stpy

. Leishmaniaes and 4-5 h

wth problemsresh growth lpful in rescu

g and don’t ures and mantle and quican using hig

stirring. Weobtaining higforces

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ained as cols). All cultivaity significan

which is essd in the daor optimal groer, if this shes medium fo

venient to gr12) in ventilace cells will adium

ase. Avoid p growth. Hsely affect su:20 on Frida

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of mid-growm at the flat

ng on solidmotile cells anre with dropd thinner (ndeprivation

tage. Also,

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s with the homedium and

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resuspend cay lyse. Centk resuspensioher speed

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ontinuous ations are tly and L.

sential for ark. After owth and elf live is r 2 more

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age much

repeated However, ubsequent ay and to es. Don’t ution. For apter 5.4

wth phase end, and

d media. nd of cells p-like cells eedle-like or other

bacterial,

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ost strain, d continue esp. after

ell pellets rifugation on of cell

fficient to ties up to

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