Why do I always get these ugly bands？ - I always end up with some fused and tandemed bands... (Dec/03/2009 )
Normally,I always end up with this kind of ugly image.
I can't figure out why.
Is there anyone can help me,please?
Why can't I get those sharp and tough bands like blow:
Yes, those bands look to good to be true, I wish I could do that too! hehehe.
AllenChiu on Dec 3 2009, 10:55 AM said:
Normally,I always end up with this kind of ugly image.
I can't figure out why.
Is there anyone can help me,please?
Why can't I get those sharp and tough bands like blow:
The center gel looks as if there is too much protein loaded in the wells. This may cause the protein to diffuse into adjacent lanes and migrate in unexpected ways.
Hi,
my guess is:
your gel is selfmade, the other gel is ordered as ready gel from a company. I don't know how they do it but it is always very sharp with commercial gels.
Stardust
Merged bands are usually due to either over-exposure or diffusion of the protein during one of the steps. Minimise the time your gels are sitting around while loading and before staining/transfer.
Your uneven bands are due to variations in the wells from when you pulled out the combs. Try pouring a stacker gel which should remove most of it, also make sure your combs are clean, wash them well (rub them to help remove stuck acrylamide) in water and with ethanol after each use.
Your smile is due to uneven heating across the gel - it is warmer in the middle so it runs faster as the pores in the gel expand. Run your gels at a lower voltage or in a cold-room with chilled buffers for better results.
lab rat on Dec 4 2009, 02:12 AM said:
seems so，thanks
bob1 on Dec 4 2009, 07:30 AM said:
Your uneven bands are due to variations in the wells from when you pulled out the combs. Try pouring a stacker gel which should remove most of it, also make sure your combs are clean, wash them well (rub them to help remove stuck acrylamide) in water and with ethanol after each use.
Your smile is due to uneven heating across the gel - it is warmer in the middle so it runs faster as the pores in the gel expand. Run your gels at a lower voltage or in a cold-room with chilled buffers for better results.
thanks for your advice，I'll try to get some ice to cool it
stardust on Dec 4 2009, 02:41 AM said:
my guess is:
your gel is selfmade, the other gel is ordered as ready gel from a company. I don't know how they do it but it is always very sharp with commercial gels.
Stardust
yes，that's mysterious