Acrylamide gel with abnormal band - (Jun/01/2007 )
Anyone has experienced this problem with acrylamide gels? The upper gel was made with 6 month old acrylamide, and the lower gel was made with one week prepared acrylamide. I know that it is a very old acrylamide to be used and I am assuming that is the origin of the problem. If so, what could be the explanation for that broad band in the gel. We were analyzing dsRNAs from plants.
Thanks
Hello
I have seen something similar with a polyacrylamide protein gel before. The cause of this was that the ethanol used on top of the resolving gel while it was setting was contaminated with protein (no idea how that happened!!).
This lead to a protein band right across the middle of the gel.
Perhaps your old batch of acrylamide has been contaminated in some way?!
I have seen something similar with a polyacrylamide protein gel before. The cause of this was that the ethanol used on top of the resolving gel while it was setting was contaminated with protein (no idea how that happened!!).
This lead to a protein band right across the middle of the gel.
Perhaps your old batch of acrylamide has been contaminated in some way?!
Yes, maybe something is growing in the arcylamide, although it looks clear to me. I should prepare smaller volumes of acrylamide and do it perhaps on a monthly basis. Thanks
we prepare large batches of acrylamide and use them until they are finished, regardless of how long it takes, and have not come across a problem like yours.
we store the acrylamide at 4C in brown glass or covered with aluminum foil.
improper storage can lead to decomposition of acrylamide to acrylic acid. the gel won't polymerize as well and can lead to broad bands.
we store the acrylamide at 4C in brown glass or covered with aluminum foil.
improper storage can lead to decomposition of acrylamide to acrylic acid. the gel won't polymerize as well and can lead to broad bands.
Thanks for the information.
We checked the reagents and all of them look fine. We always store the acrylamide batch in a brown glass at 4 C. Last January we had the same problem and thought that it was resolved preparing a fresh batch of acrylamide. Well, checking our records, it appears that this problem only happens when we run two gels in the electrophoresis chamber. The apparatus is a Mini-Protean 3 Cell by Bio-Rad. It is suppose to accommodate and run one or two gels at one time. The conditions are 100 volts for 2.5 h, constant V. We just ran one gel and it is normal, no abnormal bands. Still do not know why it happens with two gels running.