DNA extraction of animal tissue

[arm_chair27]

DNA extraction of animal tissue

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Post by arm_chair27 » Thu Jun 07, 2007 3:57 pm

i ever used Intelligene's lysis buffer. Its' protocol is very easy. you can put your sample tissue in lysis buffer -->grind it well-->incubate at 95 degree celcius for 15 min-->centrifuge-->pipette the supernatant to 95% ethanol-->centrifuge-->dry pellet and dissolve with TE buffer or DI water.
After i check DNA quality with spectrophotometry and gel electrophoresis, i's ok-->there are protein interfere a little. I would like to know what is the component of this lysis buffer. Who can guess? please tell me.thank you.

[mith]

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Post by mith » Thu Jun 07, 2007 4:17 pm

Some sort of detergent I'm guessing? In my lab we also add proteinase to it before incubating at 56C for 1-3 hrs.

[canalon]

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Post by canalon » Thu Jun 07, 2007 9:22 pm

I suspect more than one component. You can find some indication in the MSDS or safety sheets (in case you drink it you need to know what to do before you decide to extract your total DNA.

But you will probably find a strong detergent, a protein denaturant (GITC for example) and maybe something that will precipitate sugar such as CTAB.

[molecular newbie]

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Post by molecular newbie » Wed Jun 20, 2007 7:52 pm

usually we add proteinease K which denatures all the proteins....if you try adding pro K along with the lyssis buffer chances are you wont get protein interference......

[MichaelXY]

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Post by MichaelXY » Sat Jun 23, 2007 3:08 am

Geez, I had no idea what you guys were talking about, maybe after next semesters Micro Bio class, but that Pro K sounds like a good agent for jellyfish stings. Sounds better than the alternative, which I am sure you have heard of.