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DNA extraction from mouse tail

DNA extraction from mouse tail - DNA Extraction Forum

DNA extraction from mouse tail - DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.


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  #1  
Old 08-25-2008, 01:48 AM
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Default DNA extraction from mouse tail



Hi
I have started to extract DNA from mouse tail using Qiagen DNeasy tissue kit.Has anyone used this kit.My DNA concentration is in the range of 50-250ng/ul.How to increase the concentration.I use 1.2 cm of the tail and cut into pieces using scissors to start with and do an overnight lysis.
Can anyone who has been using this kit can give any helpful tips.
Thankyou.
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Old 08-25-2008, 08:25 AM
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Question Re: DNA extraction from mouse tail

Hello Bactin, Qiagen makes quite good kits however concentration of DNA can be an issue. To easily solve this simply elute with less buffer (TE or similar).

How much buffer are you eluting with now?
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Old 08-25-2008, 08:59 AM
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Default Re: DNA extraction from mouse tail

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Originally Posted by kiki06 View Post
Hello Bactin, Qiagen makes quite good kits however concentration of DNA can be an issue. To easily solve this simply elute with less buffer (TE or similar).

How much buffer are you eluting with now?
The said amount is 200ul but i have done with 100ul and now also with 50ul.with 50ul my DNA concentration was 200ng/ul.But the kit says we can extract upto 2ug or even more than that for tail genomic DNA.
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Old 08-25-2008, 11:32 AM
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Default Re: DNA extraction from mouse tail

Hi Bactin
Some questions:

Make sure the tails are submerged in the buffer completely?
After proteinase K there should be only mouse hair no tail?
Did you vortex the lysed tail before adding them to the column?
Are you using proteinase K from the kit?

also too much tail 1.2 cm may not be good as it may be less digested and hence the column will have a problem with the collagen and tissue. How much tail does the kit suggest - 0.5cm or more / less?
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Old 08-26-2008, 12:54 AM
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Default Re: DNA extraction from mouse tail

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Originally Posted by admin View Post
Hi Bactin
Some questions:

Make sure the tails are submerged in the buffer completely?
After proteinase K there should be only mouse hair no tail?
Did you vortex the lysed tail before adding them to the column?
Are you using proteinase K from the kit?

also too much tail 1.2 cm may not be good as it may be less digested and hence the column will have a problem with the collagen and tissue. How much tail does the kit suggest - 0.5cm or more / less?
the kit suggests 2 pieces of 1.2 cm mouse tail so i used the same.yes they are completely submerged as i cut them into smaller pieces before adding the buffer.after lysis there is only mouse hair (i think so) so i did a centrifugation step to remove the hair and used only the supernatant.
I vortex them well before adding to the spin coloumn and i am using the proteinase K from the kit.
yesterday with all these procedures in place the DNA concentration was 800ng/ul,the 260/280 ratio was 1.88.Is this good or can it be improved much more.
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Old 08-26-2008, 12:56 AM
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Default Re: DNA extraction from mouse tail

correction - the kit suggests 2 pieces of 0.6cm tail.
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Old 09-01-2008, 03:16 AM
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Default Re: DNA extraction from mouse tail

Hi bactin that seems much improved!

have you tried to use the DNA for PCR or other methods?

how is it going with that
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Old 12-15-2009, 06:49 PM
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Default Re: DNA extraction from mouse tail

Be sure your proteinase K is kept in a desiccant container to preserve it.
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