Hello Everyone,
I´ve been doing freshwater microalgae community profiling very succesfully, but now I´ve encountered a problem, I´ve been running some samples and recovered several bands from the DGGE without problems, but some samples (always the same ones) are not being recovered.
I´ve tried 4 times already, repeating the entire procedure, starting from the amplification of the samples.
I´m using a protocol described by Chory and Pollard (1999) for the band recovery, broadly consists on:
Elution Buffer pH 7.5 (10mM Tris-HCl, 50mM NaCl, 1 mM EDTA pH 8.0)
Overnight incubation (~25°C)
DNA precipitation (EtOH)
I´ve already changed the elution buffer and prepared new one.
The thing that strikes me the most is that some of the bands that I can´t recover look very intense and that the rest of the bands, even though they don´t look that intense, have been recovered without problems.
Anyone has ideas?
Thanks in advance!
Carolina
- Badges
- Science topic
- Similar topics
- Cell Biology
- Methods