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Trizol DNA extraction

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Leon Bach

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Apr 8, 1997, 3:00:00 AM4/8/97
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<fontfamily><param>Courier</param><smaller>We have been trying to
extract DNA from cells in culture following RNA extraction using
Trizol. We get good yields of RNA, but, after following the DNA
protocol provided with the reagent, we get a large amount of sticky
insoluble material which does not dissolve even after heating.
Absorbance at 260 nm of the supernatant suggests that there is a small
amount of DNA present.


We would appreciate any advice/suggestions.</smaller></fontfamily>

Nigel Kenward

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Apr 10, 1997, 3:00:00 AM4/10/97
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Hi,
I found that I had to do double the number of washes in the variuos
solutions to get the DNA really clean. There was loads of DNA then (tho'
it got a bit sheared).

Has anyone any idea if it's all genomic DNA or if transfected plasmids
replicating independantly from the genome will be purified too. I
haven't detected any plassmid but an unhelpful lady on the technical
support reckoned all DNA would be purified (in a manner that suggested
that 5 yr old knew that !!)

Cheers,
Nige
--
____ ___
|\ | | / \ / | NIGEL KENWARD,
| \ | | | ___ |__ | Dept. Biochemistry,
| \ | | | | | | Nottingham University Medical
School.
| \| | \____/ \___ |___ Clifton Boulevard.
------------------------------- Nottingham. NG7 2UH.
Tel (0115 924 9924) ext 44789 Fax (0115 942 2225)
email:mbx...@vax.nott.ac.uk "You can tell me.......I'm a doctor"

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