
Western Bizarre
#1
Posted 30 January 2010 - 04:38 AM
As you can see there must be some thing wrong with the SDS loading buffer directly harvested samples.They are twisted and partially diffused(in the middle).
Someone told me that it seems the SDS in 1s samples are not enough...
So what do you think ?
Thanks a lot
#2
Posted 31 January 2010 - 03:33 PM
#3
Posted 02 February 2010 - 02:13 AM
...but I'm afraid that the odd that every #1 lanes have uneven bottom while #2 lanes are not is very slim.Gel images taken of several gels using the same combs for set up is my guess... the wavy band is probably due to uneven bottom of the well, this is usually the result of having a damaged or dirty comb.
#4
Posted 02 February 2010 - 09:06 AM
talent does what it can
genius does what it must
i used to do what i got paid to do
#5
Posted 03 February 2010 - 10:00 PM
Thank you..but how could anyone possibly exclude the salt that you mention while harvest the cells?as you know there seems to be heavier salt in PBS,if you wash cells before harvesting.was there any medium left behind prior to harvest of sample 1? residual salt from the medium could cause the distortion you are seeing.
#6
Posted 04 February 2010 - 01:37 AM
I also started to think that the problem is with the buffer. SDS might be a good suggestion. If you solve the problem, could you please poste it

#7
Posted 04 February 2010 - 09:15 AM
I got a plan now,which has nothing to do with SDS.I have a similar problem (see the post Diffused bands few posts below yours) and I also use 1× loading buffer. I did not compare different samples though.
I also started to think that the problem is with the buffer. SDS might be a good suggestion. If you solve the problem, could you please poste it
tell me if you want to hear it

#8
Posted 04 February 2010 - 01:03 PM
just explaining why you were seeing what you were seeing, not how to resolve the situation.Thank you..but how could anyone possibly exclude the salt that you mention while harvest the cells?as you know there seems to be heavier salt in PBS,if you wash cells before harvesting.was there any medium left behind prior to harvest of sample 1? residual salt from the medium could cause the distortion you are seeing.
by the way, the distortion did not appear to be bad enough to worry about. if it is then you will need to find a way to remove the contaminating salt.
talent does what it can
genius does what it must
i used to do what i got paid to do
#9
Posted 05 February 2010 - 06:42 AM
I got a plan now,which has nothing to do with SDS.I have a similar problem (see the post Diffused bands few posts below yours) and I also use 1× loading buffer. I did not compare different samples though.
I also started to think that the problem is with the buffer. SDS might be a good suggestion. If you solve the problem, could you please poste it
tell me if you want to hear it
yes, of course, please tell me your plan

#10
Posted 22 February 2010 - 03:17 AM
sorry,my plan has failedI got a plan now,which has nothing to do with SDS.I have a similar problem (see the post Diffused bands few posts below yours) and I also use 1× loading buffer. I did not compare different samples though.
I also started to think that the problem is with the buffer. SDS might be a good suggestion. If you solve the problem, could you please poste it
tell me if you want to hear it
yes, of course, please tell me your plan
