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Storing mouse tails for eventual genotyping...


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#1 assembler01

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Posted 07 May 2010 - 08:57 AM

If you have a bunch of tails you clipped or someone just gave you on Friday, what is the best way to store them until Monday? I normally use the qiagen puregene kit for genomic DNA isolation and digest them overnight. Can I leave the tails in the cell lysis solution over the weekend at 55 degrees? Should I leave them whole in the refrigerator? Would freezing them preserve them better or would it degrade the DNA?

I saw a protocol for sending them through the mail on a company's web site that said you can store or ship them in 70% ethanol at room temperature (or I imagine 4 degrees as well) but I'm curious if that would interfere with the puregene protocol.

I'm sure someone out there has run into this before. What do you normally do and do you notice degradation of DNA amount or quality with weekend storage?

Thanks a lot.

#2 shldbcrzy

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Posted 07 May 2010 - 10:50 AM

I have used the same system and if i had them over the weekend i would just freeze the tails immediately till further use.

#3 macroman

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Posted 07 May 2010 - 12:58 PM

I do incubate tail snips at -20C as routine atleast for overnight.

Hope this solves yr purpose

#4 zodiac1505

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Posted 01 July 2010 - 06:38 AM

I also do the same. I just chuck them at -20 till I have time to do the tail lysis and DNA extraction :(

#5 pmaj

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Posted 01 July 2010 - 12:12 PM

I used to extract genomic DNA via the crude method and store it in TE buffer. Here is the protocol I use:
1. Each tail should be in a clean eppendorf tube.
2. Add 500Ál of tail lysis buffer containing Proteinase K (PK) to each tube.
3. Incubate tail samples in 50-60C water bath overnight.
4. Add 250Ál saturated (6M) NaCl to each tube.
5. Shake tubes vigorously (~ 20 times) and incubate tubes on ice for 10 minutes.
6. Spin tubes on low speed (#6 on Hemle centrifuge) at 4C for 10 minutes.
7. Remove supernatant and place into a clean eppendorf.
8. Add 650Ál isopropanol and invert to mix. Incubate tubes at room temperature for 15 minutes.
9. recover DNA by centrifuging, max speed, 10 minutes at room temp.
10. Place tubes inverted on bench and allow to air dry 5 minutes.
11. Add 200Ál of TE pH 7.5 or sterile water to each tube. Incubate in 50-60C water bath for * 10 minutes. Resuspend pellet by pipetting up and down several times.

I have stored my samples in the TE buffer for a couple of weeks at 4 deg C, but the DNA does degrade over a period of time, but it should be fine for a week or two at 4 deg C in TE buffer for genotyping.

Good Luck

Pmaj

Edited by pmaj, 01 July 2010 - 12:13 PM.

There is a fine line between persistence and obstinacy. I have come to realize the key is to choose a problem that is worth persistent effort.
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