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indoubt's Content

There have been 40 items by indoubt (Search limited from 18-September 18)



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#13275 Primary and diploid cultures.

Posted by indoubt on 31 March 2005 - 12:18 AM in Cell Biology

What are the difference between Primary and diploid cultures?


Thanks.



#13235 100X=100%?

Posted by indoubt on 30 March 2005 - 06:44 AM in General Lab Techniques

Triton-100X is the same as 100%. Does it also mean that if a solution is 50X then it is 50%? How come?


THanks



#13231 96 and 24 well plate

Posted by indoubt on 30 March 2005 - 06:06 AM in Cell Biology

hi
volume of a 96well is 150ĩl
the difference is the diameter of the well. in a 24well diameter  is about 4fold greater.

<{POST_SNAPBACK}>



So it is 150ul and not 150ml? If the well of a 24 well plate has greater diameter then why it has less volume ? <_<


Thanks.



#13217 96 and 24 well plate

Posted by indoubt on 30 March 2005 - 03:32 AM in Cell Biology

What is the difference between a 96 and 24 well plate? Why the volume in a well (about 150ml) of a 96 well plate is larger than a well (2ml) of a 24 well plate ? <_<



Thanks.



#13017 Incubate suspension cells?

Posted by indoubt on 25 March 2005 - 11:57 AM in Cell Biology

How can i incubate suspension cells? Should they be stirred all time or should i just put them in the 5% incubator like anchorage-dependent cells?

How can i feed suspension cells?


Are the cells from the hypophysis tissue cells or neural cells? And do the general cell culture rules also apply to the neural cells?



Thanks.



#12983 Plasmids in eluting buffer

Posted by indoubt on 24 March 2005 - 08:38 PM in Molecular Biology

Mini/maxi prep of plasmids are stored in the elution buffer. I wonder when i use these plasmids for further working like sequencing or PCR should i first purify them from this eluting buffer or can i use them directly in this buffer? :o



Thanks.



#12812 Ideas about these questions, please.

Posted by indoubt on 23 March 2005 - 03:20 AM in Cell Biology

hi

poly a sequence upstream a reporter gene is for ensuring the good RNA termination, and translation.

<{POST_SNAPBACK}>


Usually the poly A sequence lays downstream of a gene to give it good RNA termination and translation. BUt when it lies upstream is something new . Upstream means before the promoter. How come? :(



#12776 Ideas about these questions, please.

Posted by indoubt on 22 March 2005 - 06:36 PM in Cell Biology

Ideas about these questions, please.

#Why is Poly-A sequence upstream of a reporter gene?
#Isogenic cell lines?
#Null cell lines?

Thanks. :P



#12608 pH and osmolarity?

Posted by indoubt on 19 March 2005 - 03:15 AM in Cell Biology

Should pH always be between 7.0 to 7.4 in the medium? What about the osmolarity? :unsure:

Thanks.



#12579 passage number in mammalian cells

Posted by indoubt on 18 March 2005 - 03:02 AM in Cell Biology

be careful. there is no law to all this (cell lines themselves are anarchists)...

of course immortal cell lines can change their character. they donīt do it that easily. it depends... on the growth conditions/handling, the cell line etc.

sorry itīs not that easy.

Are all cancer cells immortal cells? Or are there only cells that express an immortal protein that become immortal cells whether they are normal cells or cancer cells? :D

Thanks.



#12492 good links of Microarrays?

Posted by indoubt on 17 March 2005 - 03:55 AM in General Lab Techniques

Lemme know what kind of info you need, Protocols? Just basic Info? Advanced Data Analysis stuff? I have viewed and used info from a lot and I might be able to give you some links.

Phantom.

This is the first time i will work with this technique, but i can't find any info or good links about it. I would like to know about it as much as i can. I would love to have them all. This would help me alot in my experiments. Thank you very much. :rolleyes:



#12426 RT- PCR?

Posted by indoubt on 16 March 2005 - 07:39 AM in Molecular Biology

links and info of RT- PCR?

appreciate for any inputs. :D



#12401 good links of Microarrays?

Posted by indoubt on 16 March 2005 - 04:28 AM in General Lab Techniques

anyone with good links of Microarrays?


thank you.



#12400 passage number in mammalian cells

Posted by indoubt on 16 March 2005 - 04:25 AM in Cell Biology

thanks.



#12391 freezing of cells

Posted by indoubt on 16 March 2005 - 02:39 AM in Cell Biology

Most protocols say to freeze cells at 3*10 to the 7 cells per mL, this is standard across most cell lines and cell types.

you meant 3x10^7 cells/ml?



#12390 passage number in mammalian cells

Posted by indoubt on 16 March 2005 - 02:20 AM in Cell Biology

Thanks.

So immortal cell does not die and does not change character, while mortal cell dies and changes character when passage number is high? :D



#12351 passage number in mammalian cells

Posted by indoubt on 15 March 2005 - 02:59 PM in Cell Biology

well, officially you got 6 passages left...counting down...3...2...1 - pay another 400 bucks for a fresh vial from your favourite supplier...

so after my frozen stock has reached the last passage number then i should get a new vial from the supplier? what if i freeze down the last passage number and keep thaw and passage it? will my results get affected?

ever since i started working with cell lines, i have been wondering if people do it that way in real world.

what do you mean? :D



#12343 passage number in mammalian cells

Posted by indoubt on 15 March 2005 - 01:27 PM in Cell Biology

if a cell line has a maximum of 10 passages before it changes character and i only have done 4 passages of it and made the frozen stock of some of the cells. how many passages does this frozen stock have if i thaw it and passage it? will it be only 6 passages left or 10 passages?

thanks for any inputs.



#12318 freezing of cells

Posted by indoubt on 15 March 2005 - 03:39 AM in Cell Biology

:unsure: thanks.



#12308 freezing of cells

Posted by indoubt on 15 March 2005 - 02:34 AM in Cell Biology

So in order to prevent a passage of cells when thawing them, and by the way damage half of the quantity, passage is done before freezing.


What do you mean with passage? Can you explain what you mean? Thank you very much.



#12258 freezing of cells

Posted by indoubt on 14 March 2005 - 03:02 PM in Cell Biology

my protocol says this:

1. spin down the remaining cells.

2. resuspend cells with 1.5ml medium.

3. add 1.5ml 2X freezing medium.

4. freeze down cells in 1.6ml cryovials.

my question; why should i add 1.5ml medium (step 2) when my vial only has a total volume of 1.6ml. should i ignore step 2? any ideas? B)

thanks.



#12119 right medium and serum?

Posted by indoubt on 11 March 2005 - 05:40 AM in Cell Biology

is there a website which shows which medium and serum is for a cell line? i have the problem looking for the right medium and serum.

thank you.



#11654 micro-matrixes

Posted by indoubt on 02 March 2005 - 04:40 AM in Molecular Biology

anyone has good links about protein and DNA micro-matrixes?

thanks. B)



#11549 .....

Posted by indoubt on 28 February 2005 - 11:22 PM in Microbiology

.....



#11490 measuring protein concentration

Posted by indoubt on 28 February 2005 - 01:34 AM in Protein and Proteomics

thanks, mike!

but what happens if i add different volume of dyes into the samples and standard solutions? should i in this case use the dye volume in my calculation too? :huh:




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