How to concentrate DNA
Posted 05 July 2012 - 04:05 AM
I would like to concentrate my DNA sample.
At this moment, i'm trying to maximize my DNA sample during extraction, by using low elution volumes and a higher amount of input material.
By doing this my concentration enhanced from 68 ng/ul to 156 ng/ul.
Is there an additional solution to enhance my DNA concentration besides lower elution volumes?
Posted 05 July 2012 - 07:39 AM
2. Holding elution buffer 1-2 minutes before centrifugation (if this step required like in miniprep) is also recommended.
Posted 05 July 2012 - 07:48 AM
These have 30kdalton and 100kDalton cuttoffs but they also do other cuttoffs depending on how long your DNA is that you want to retain.
Here's the brochure:
I used to use their predecessor, the Microcon, to great effect and I don't suppose the Amicon is any less capable. They're not inexpensive but they can concentrate DNA well and also help get rid of salts and other contaminants.
Posted 09 July 2012 - 11:09 PM
Posted 09 July 2012 - 11:18 PM
so after eluating DNA, i just have to add ethanol/isopropanol to the eluaat? How many?
Do you have a protocol for this?
Posted 09 July 2012 - 11:25 PM
1. Add equal volume of chilled isopropanol, mix and incubate at -20 C for 30 min
2. Pellet down the DNA by centrifuging at 10000 g / 10 min / 4 C
3. If you have used elution buffer (usually TE) for elution, give 70% ethanol was to the pellet (add 1 ml 70% ethanol, incubate at RT for 5 min, spin 10000 g / 10 min / RT). If you have used water for elution, no need to give 70% ethanol wash
4. Dry the pellet (after complete removal of the supernetant using pipette) at RT for 10 min
5. dissolve in the desired volume of water/buffer
Posted 11 July 2012 - 06:04 AM
Edited by mdfenko, 11 July 2012 - 06:05 AM.
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