|
Nexterion
Spot
For
the spotting and binding of DNA-probes onto Nexterion Slide E and
Nexterion Slide AL we recommend the optimized spotting buffers
Nexterion™ Spot. Based on phosphate buffer with an alkaline pH,
they ensure efficient immobilization and homogeneous spots.
As an alternative 3x SSC or 1.5 M betaine in 3x SSC can be used. The
latter is especially useful to avoid evaporation of the spotting
solution during the spotting process.
Nexterion
Slide A+ is compatible with most common spotting solutions. The
betaine or DMSO containing spotting solutions are especially
suitable when probe evaporation during long spotting runs is
expected.
Recommended
spotting solutions for Nexterion Slide A+
|
Spotting
Solution |
Remark |
|
50%
DMSO |
Larger
spot size, prevents evaporation problems during long spotting
runs |
|
3xSSC |
Smaller
spots, standard aqueous spotting solution |
|
3xSSC
+ 1.5 M betaine |
Larger
spots, prevents evaporation problems during long spotting runs,
very homogeneous spots |
|
Nexterion
Spot |
Smaller
spots, phosphate buffer based solution |
FBNC buffer
The
FBNC buffer composition contains 0.5 M betaine, 25% (v/v) formamide
and 0.5 µg/µl nitrocellulose.
Of
the combinations surveyed by Gunnar Wrobel et
al.(2003), combining Nexterion Slide E and FBNC as the spotting
buffer seems to be the best compromise in terms of spot diameter,
signal intensity and sensitivity. This configuration allowed a
density of up to 70 000 spots per slide.
Bibliography
reference :
Gunnar
Wrobel et
al. , 2003 - Nucleic Acids Research, 2003, Vol. 31, No. 12 e67
Optimization
of high-density cDNA-microarray protocols by ‘design of
experiments’ |
