I am following a protocol that requires the stacking buffer be adjusted to a pH of 6.8. I accidentally overadjusted it to around 6.1 so I used some NaOH to compensate the pH back to 6.8. Would this affect the stacking gel performance or not really?
0
3 replies to this topic
#2
bob1
-
- Global Moderators
-
- 4,350 posts
Thelymitra pulchella
224
Excellent
Excellent
Posted 14 August 2012 - 01:13 PM
The NaOH adds Na to your solution (the OH- combines with the H+ in the solution to form water), if the assay is dependent on salt you may have a problem, but it should be fine most of the time.
The usual way to do this sort of thing is to make a tris solution of pH 6.8 and add that to your stacking buffer.
The usual way to do this sort of thing is to make a tris solution of pH 6.8 and add that to your stacking buffer.
#3
mdfenko
-
- Active Members
-
- 2,247 posts
an elder
78
Excellent
Excellent
Posted 15 August 2012 - 05:07 AM
the addition of naoh will affect the stacking buffer.
if you overshoot then you should add more basic component (tris), calculated for a greater volume, readjust pH, then bring to the new final volume.
if you overshoot then you should add more basic component (tris), calculated for a greater volume, readjust pH, then bring to the new final volume.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#4
pinkc
-
- Active Members
-
- 5 posts
member
0
Neutral
Neutral
Posted 16 August 2012 - 11:16 AM
Ack! OK thanks a lot for replying - I haven't used it yet but now I will dump it out and remake it and be careful about adjusting pH.
