1. SCHOTT Jenaer Glas GmbH
Technical Support
Phosphate Buffer based Spotting Solutions
Slide E Slide A Slide
A+
Slide H Slide
AL
Comments
Nexterion Spot (phosphate buffer based)
 
   add detergent to
increase spot
size1
50 to 300 mM Phosphate Buffer, pH 8.0
to 9.0      50 mM
phosphate
buffer =
CodeLink Buffer
300 mM Phosphate Buffer + 0.005%
Tween20 + 0.001% sarcosyl      Spot H (50)
150 mM Phosphate Buffer + 0.001%
Tween20      Spot H (30)
1
detergents: SDS, sarcosyl, Tween, Triton, Cetyl-trimethylammoniumbromid, final concentrations
needs to be determined empirically, usual f.c. range between 0.001% and 0.05%

2. SCHOTT Jenaer Glas GmbH
Technical Support
Low Evaporation Spotting Buffers
Slide E Slide A Slide
A+
Slide H Slide
AL
25% to 50% ethylene glycol in
phosphate buffer
 ? - ? -
25% to 50% ethylene glycol in H2O
- ? - ? 
25% to 50% DMSO in H2O
-   - -
25% to 50% DMSO in phosphate buffer
- ? ? 2 ?
3x SSC + 1.5 M betaine
 
 

2
used by a customer

3. SCHOTT Jenaer Glas GmbH
Technical Support
DMSO containing Spotting Buffers on
Slide E
Spot Size
SMP3
Spot Size aQu
K2801
25% DMSO, 1.5x SSC, 0.005% SDS* 90 µm 160 µm
30% DMSO, 1.5x SSC 70 µm 90 µm
50% DMSO, 1.5x SSC 70 µm - (inhomogeneous
spots)
50% DMSO, 1.5x SSC, 0.005% SDS* 60-70 µm 110 µm
50(30) % DMSO, 2x or 2.5x SSC 100 to 130 µm
*
higher concentrations of SDS may lead to precipitation, other detergents also work: Triton X100,
cetyl- trimethylammonium bromide, both with a f.c. of 0.025%)
Problem: with DMSO as spotting solution for Slide E are small spot sizes (as low as 50 µm)
Solution: add SSC or SSC plus detergent, do not add phosphate buffer or Next Spot

4. SCHOTT Jenaer Glas GmbH
Technical Support
Adjust Spot Size for DMSO containing
Spotting Solutions
data shown on previous slide are based on a final concentration of 10 µM
(70mer, Operon)
if a customer has Oligos already dissolved in DMSO, usually the final
concentrations of oligos is 20 µM and higher
to adjust spot size, SSC and SDS may be added:
Example: customer has 50 µl of 20 µM oligo dissolved in 50% DMSO
add: 50 µl of 50% DMSO containing 3x SSC and 0.01% SDS
final concentration of components 10 µM oligo (sufficient for Slide E)
50% DMSO
1.5x SSC
0.005% SDS

5. SCHOTT Jenaer Glas GmbH
Technical Support
Slide E Slide A Slide
A+
Slide H Slide
AL
Comments
3x SSC     
3x SSC + 1.5 M betaine  
   low evaporation
50% DMSO in 0.5x SSC* 3
? ? ? ? low evaporation
SSC based Spotting Buffers
3
used by a customer, most probably will work also on Slide A, A+
* for other combinations of DMSO and SSC see also slide “DMSO containing spotting buffers on
Slide E”

6. SCHOTT Jenaer Glas GmbH
Technical Support
Slide E Slide A Slide
A+
Slide H Slide
AL
Comments
3x SSC     
3x SSC + 1.5 M betaine  
   low evaporation
50% DMSO in 0.5x SSC* 3
? ? ? ? low evaporation
SSC based Spotting Buffers
3
used by a customer, most probably will work also on Slide A, A+
* for other combinations of DMSO and SSC see also slide “DMSO containing spotting buffers on
Slide E”