composicion de la solucion tl- hepes como extensor

4/15/2015 TLHepes

http://www.ansci.wisc.edu/jjp1/ansci_repro/lab/procedures/media/tl_hepes.html 1/1

TLHEPES

Component M.W. mM amount/100mlamount/500

ml1.CaCl2.2H2O 147.02 2.0 30.0mg 150mg2.KCl 74.56 3.2 23.5mg 118mg3.MgCl2.6H2O 203.3 0.5 10mg 50mg4.NaH2PO4.H2O 137.99 0.4 5.6mg 28mg5.LacticAcid(Sodiumsalt60%w/wsyrup)

112.1 10.0 186mg(141l)

930mg(705l)

6.NaCl 58.44 114.0 666mg 3330mg7.NaHCO3 84.01 2.0 16.8mg 84mg8.HEPES(FreeAcid) 238.3 10.0 238mg 1190mg9.NaPyruvate(fromstock) 110.0 0.2 1.0ml 5.0ml

10.BSA(FractionV) 3mg/ml 300mg 1500mg11.Gentamycin(fromstock) 50g/ml 100l 500l

Addapproximately80%ofthedoubledistilledwaterneedtoabeaker.Withstirring,addcomponents18inorder.AdjustpHto7.4at2022Cusing1or2NNaOH.Addwatertobringto100%ofvolume.Thiscanbestoreduptotwoweeksasastockat5Ciffilteredandkeptsterile.Generallywemakeup500mlatatime.Ondayofuse,addcomponents9and10.Notethatthesecanbeforanyvolumedesiredforusethatdayandneednotbe100or500ml.Filtertosterilizewith0.2mfilters.SeeStocksolutionsforcomponents9and11.

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