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Seeding plates


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21 replies to this topic

#16 pito

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Posted 11 May 2011 - 06:54 AM

??????????????????
I am blank


are you sure your info is right?

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#17 casandra

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Posted 11 May 2011 - 06:56 AM

??????????????????
I am blank

What do you mean blank? :) Btw, are you and kumara one and the same? Will you double-check again the number of cells you need bec 6.25 million per well seems pretty steep (perhaps it's one too many zeroes?)...anyways, just do a rough calculation....you need about 75 million cells i.e. 2.5 mill/ml X 30 ml (2.5 ml X 12 wells) or 6.25 mill X 12 wells...and you only have about 5 million cells in total (240,000 cells X 20 ml) kumara did write 20 ml or like pito wrote only 2.4 million if you only have 10 ml...are you sure that your stock is only this much (one or two less zeroes)? You may have to expand them more if you really need the 6.25 million per well...

Edited by casandra, 11 May 2011 - 07:06 AM.

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- hobglobin, personal comment about my beauteous photo......

#18 harry348

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Posted 11 May 2011 - 07:07 AM

I resuspended the pellet in 20 ml medium and in this i got around 24*10000 as the number of cells per ml

#19 casandra

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Posted 11 May 2011 - 07:12 AM

I resuspended the pellet in 20 ml medium and in this i got around 24*10000 as the number of cells per ml

so you only have 4.8 million cells in total......therefore, if your numbers are all correct, you don't even have enough cells for one well.....
"Oh what a beauteousness!"
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#20 pito

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Posted 11 May 2011 - 07:22 AM

If I remember correctly, you once wrote 625.000 cells per well in another post not 6mil....

(use . between your numbers, its easier to read and you would make less mistaks, because now I have to idea you made typing errors in the numbers and you dont even know anymore what it has to be...)

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#21 leelee

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Posted 11 May 2011 - 08:42 PM

Mate, as cassandra said, even if it is a well of a six well, 6 million cells is FAR too many!!! A confluent well of a six well will only have just over 1 million cells.

If you add 6 million cells to your wells (which you can't because you don't have enough cells anyway), they will be so over crowded that it would be extremely bad practice to use them for any kind of experiment.

Perhaps you should double check your numbers and explain again from the beginning?
It would help us all if you used correct scientific notation for your numbers too. It is far too easy to type in an extra zero and cause confusion.

#22 harry348

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Posted 12 May 2011 - 01:13 AM

ok Mate

Today im doing the cell splitting and seeding and i wil let you know what my count is




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