Immerse the formalin-fixed samples in TE buffer for at least 2 hours or longer (e.g. overnight) before starting the DNA extraction? What is the purpose of it? What TE buffer will do?

There is a kit available from Zymo Research which can be used very efficiently. It works well for us.

Try the phenol chloroform extraction, I have done it before and had success. I also have known people to have success with the qiagen spin columns.

Good luck.

I don't know if your ever tried it but the MasterPure™ DNA Purification Kit from epicentre seems to work according to my collegues.... so it could be a solution

http://www.epibio.com/products/nucleic-acid-purification-extraction-kits/dna-purification-genomic/masterpure-complete-dna-and-rna-purification-kit?protocols

hope it will help.

Noris

Try the BlackPREP FFPE DNA Kit (Analityk Jena/Biometra), extraction is performed without dewaxing.

Good luck

This paper uses a method to extract DNA from formalin fixed/ wax embedded tissue. http://www.ncbi.nlm.nih.gov/pubmed/16049299

You can also refer to :

http://www.protocol-online.org/prot/Protocols/DNA-Extraction-from-Archival-Formalin-fixed--Paraffin-embedded-Tissue-Sections-3159.html

Using most of commercial kits, incubation with proteinase K is at 56ºC for 1-2h. Try to incubate instead over night (at 56ºC). Add 10 ul more of proteinase K next morning and incubate for an additional hour at 56ºC (all times with shaking). Thereafter follow the recommended protocol. So you should be able to obtain DNA for PCR from an even very small sample. DNA extracted from wax embedded tissue may have a very bad quality, note to amplify small fragments (no more than 250 bp).

You can use the DNeay blood & Tissue kit from Qiagen, there is also a pretreatment for praffin-embedded tissue. se manula Deasy Blod & Tissue handbook

Both ChargeSwitch gDNA (Invitrogen) and QIAamp DNA mini kit (Qiagen) works well for FFPE samples.

We do not have problems, using the methodology from this paper.

Good luck!

Stefanoff CG, Hassan R, Gonzalez AC, Andrade LA, Tabak DG, Romano S, Zalcberg

IR. Laboratory strategies for efficient handling of paraffin-embedded tissues for

molecular detection of clonality in non-hodgkin lymphomas. Diagn Mol Pathol. 2003

Jun;12(2):79-87.

I always use Qiagen Micro Amp kit. One thing I do is extend the protease incubation overnight. Always get way better quantity and purity than with a 3 hour incubation. Also do your self a favour and remove most of the wax before the xylene step. This improves 260/230 ratio.

I have some experience with wax process tissues , when it is already in section, even after staining.

1 you can just scratch the tissue out in an eppendorf,

2 use the lysis buffer you use for tail.

3 pass it through a column like quiagen gel extraction

4 run your PCR on this material

genotypage works fine ;)

cheers

Please find attached and article for the same.

Thanks to everyone for thier suggestions

Pleae see attached article.

Immerse the formalin-fixed samples in TE buffer for at least 2 hours or longer (e.g. overnight) before starting the DNA extraction? What is the purpose of it? What TE buffer will do?