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sheath buffer preparation


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5 replies to this topic

#1 ujla80

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Posted 16 January 2010 - 08:54 AM

hi all :o

i m making stock sheath buffer(10X) for FACS but not getting clear solution. i am adding
EDTA =1.8 gm
KCl = 1.25
LiCl3 = 2.15
KH2PO4 = 1.3
Na2HPO4 = 11.75
NaCl = 40.6
NaOH = 0.5
in 500ml DDW.
solution becomes milky after adding Na2HPO4 (which i add in the end). what could be the reason?
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#2 pDNA

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Posted 16 January 2010 - 10:10 AM

what is your pH? Most of the time its an issue of pH ...try to lower the pH (around 2) than add the rest and readjust the pH slowly.

Could be that you added the Na2HPO4 to fast? ...try it more slowly.

Regards,
p

#3 ujla80

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Posted 17 January 2010 - 05:42 PM

thanks P :rolleyes:

actually now i know where the problem was...constituents are to be added in order...i added NaOH after adding EDTA which should be added in the end...this was resulting into turbidity after adding Na2HPO4.

regards,
ujla

but still i want to know why NaOH is causing problem??










what is your pH? Most of the time its an issue of pH ...try to lower the pH (around 2) than add the rest and readjust the pH slowly.

Could be that you added the Na2HPO4 to fast? ...try it more slowly.

Regards,
p


god is great

#4 mdfenko

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Posted 19 January 2010 - 07:20 PM

it's not the naoh that is causing the problem, it is the edta. edta will not go into solution if the pH is low.
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#5 ujla80

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Posted 23 January 2010 - 05:17 AM

hi

yes EDTA gets dissolved at ph 8 but i added naoh after adding edta (out of order) so that it gets dissolved, as a result, solution could not be prepared..it was milky.

it's not the naoh that is causing the problem, it is the edta. edta will not go into solution if the pH is low.


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#6 mdfenko

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Posted 25 January 2010 - 07:13 AM

how old is your naoh (did you use pellets? they get coated with carbonate by absorbing co2 from the air)?

you may have introduced an insoluble carbonate into your solution.
talent does what it can
genius does what it must
i do what i get paid to do




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