Hello
I am facing problem in swelling the ssDNA cellulose resin in water or Tris-EDTA buffer (10mM Tris pH8.0 and 1mM EDTA). Even after 72 hours it is still slimy/mucoid powder and is blocking the flow when packed in a column.
There is no protocol given on any website for the usage. Can somebody kindly help
regarding the same?
found that adding liquid to dry gels/resins created lumpiness but sprinkling the gel onto the liquid seemed to lead to more uniform swelling
Adding some salt might help. You can wash it off again after the column is packed. Be very careful when handling the cellulose not to stir it around too much or it will break up into fines that will clog the column.
Three suggestions:
1. Change the molar salt concentration (Trial and error)
2. Use a relatively lower pH(around 7) for swelling of the gel and then equilibrate with the buffer of choice in column the gel in column may swell a little after this..
3. Degas the swollen gel briefly before loading the column and load the column carefully avoiding any accumulation of bubble.
Sorry for the late response. I hope I could see this earlier. I had the same problem. I called the tech support and they replied: this is a 'feature' of the resin. There are many 'fines' with the resin, which really blocks the column. My problem was not so severe since flow could still went through the column but resulted in high pressure. You should remove supernatant (with fines) and change to fresh buffer, re-suspend, let it settle down and remove any additional fines. You probably need to repeat this 3-8 times to get rid of all fines.
- Badges
- Science method
- Similar topics
- Chemistry
- Biochemistry
- Biochemical Methods
- Protein Purification Techniques