Storing mouse tails for eventual genotyping...
Posted 07 May 2010 - 08:57 AM
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.
Posted 07 May 2010 - 10:50 AM
Posted 07 May 2010 - 12:58 PM
Hope this solves yr purpose
Posted 01 July 2010 - 06:38 AM
Posted 01 July 2010 - 12:12 PM
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.
Edited by pmaj, 01 July 2010 - 12:13 PM.