Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.
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Chelex is great for DNA extraction for difficult samples often seen in forensic type work and inexpensive. This is from a Biorad leaflet..
Chelex 100 molecular biology grade resin is a highly pure, pipettable, nuclease and ligase inhibitor-free chelating resin specifically designed and certified for extraction of PCR-ready template DNA. Chelex 100 molecular biology grade resin accommodates the stringency required for a PCR-quality product by ensuring the complete removal of PCR inhibitors (contaminating metal ions that catalyze the digestion of DNA)...
Its a size thing as the resin has many applications and most people use the larger 200-400 mesh for DNA work. I am sure someone out there has got the real reasons but in practical terms go for the larger one.
source: BW Parky
I have a problem with chelex too. The difficulty lies in the lack of dissolve. I've been trying everything: boiling water, tris + 4NaOH (pH 11 and even more in the end), tris with acetic acid... and nothing. All this method i found in protocols located in Internet. unfortunatelly all publications i've found, wich include chelex extraction, didn't mention about dissolving it.
I am possessing chelex 100 in a form of tiny wihite crystals. if anybody know about solution of my problem, please, help.
Ps. sorry for my english
The chelex 100 solution is not going to dissolve in its de-ionized water. I have used chelex for quite some time, and I have found that the best extraction results are as follows:
1)spin down a sample to a pellet in a 1.5ml microcentrifuge tube.
3)Add 100 micro liters of 5% chelex 100 (shake first to get the solution to mix, and pipette quickly into your sample before the chelex settles to the bottom.)
4)boil in water for 10 minutes
5)spin down at13,000rpm for 2 minutes (must be this fast or the DNA final content will be much less.)
6)pipette off the supernatant (The DNA is in the supernatant) and pipette into a new 1.5 ml micro tube. If the supernatant is cloudy, or concentration readings are not accurate, leave the sample in 4degree overnight, and spin down again at 13,000rpm and take the supernatant off again to a new tube.
-Hope this helps
7 posts • Page 1 of 1
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