Who can help me?
Posted 25 April 2002 - 06:33 AM
Posted 26 April 2002 - 02:39 AM
did you perform a proteinase K digestion (55°C ON) before pheno/chloro extraction? Your product looks like large amount of proteins contamination that you can not remove only by solvants extraction. Maybe it will be better.
Posted 26 April 2002 - 03:44 PM
I have added PK in the lysis buffer ( final concentration is 150ng/ml)and incubated them overnight at 55C.I think it is enough.
Posted 26 April 2002 - 04:11 PM
Posted 02 May 2002 - 03:20 AM
i use to extract genomic DNA from different source by following this procedure:
- lysis and RNase A treatment 2 h @ 37°C (vol=700µl ie)
- add 30 µl PK 10mg/ml and incubate 55°C ON.
- phenol/chloro extraction 1h @ 4°C under soft rotation.
- chloro/isoamyl alcool (24:1)
- 1 Vol Isopropanol precipitation.
Maybe I use a higher conc of proteinase K than you...I don't know, but I think it is a protein contamination.
Posted 02 May 2002 - 03:39 AM
Posted 20 May 2002 - 01:56 PM