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Until this gel has finished running?
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Due to this fact, the temperature should be generally as small as possible until the gel point is reached to reduce the diffusion, without considering the higher crosslinking speed with higher temperatures.
Therefore, the applied vacuum should be as high as possible (Eq. 11) to reduce the amount of side reactions until the gel point is reached.
After 3 h of stirring, 6.6 mL tetraethylorthosilicate (TEOS) was added drop wise and the stirring was continued until the gel became homogen.
To create the gel blocks, a mixture of tap water and 2% agarose gel powder (Agarose A9539, Sigma-Aldrich Corporation, St . Louis MO, USA) was heated in a microwave oven until the gel powder was completely dissolved.
Based on the fact, that the diffusion is possible until the gel point is reached, the correlating gel time should be minimized (Eq. 9) to reduce the amount of diffused contaminations, resulting in a smaller contaminated resin volume.
Application of Carisolv™ gel continued until the gel stayed clear.
Put the culture dishes under 4°C for 15 min until the gel was solidified.
Place in the first set of plates and repeat until the gel casting chamber is full.
The glass plate with the freshly coated gel solution was put into an oven at 80 °C until the gel was dry (approximately 10 min).
Protein spots were excised manually from Coomassie blue-stained gels and washed twice in 25 nM NH4HCO3 for 10 minutes until the gel pieces become transparent.
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