PCR bands - why not straight ?&%#@&& - (Oct/14/2008 )
I have to clone a PCR product ~1 kb into a cloning vector.
And the PCR is coming in fact a good amplification...but around the single band there is a little glowing...what could be that.
apart from that can anyone tell why sometimes the band are not straight but slightly curved upwards from the ends. Even the marker ran along with it is curved. the gel was runned on 1% agarose.
beulah
-beulah-
QUOTE (beulah @ Oct 14 2008, 04:50 AM)
I have to clone a PCR product ~1 kb into a cloning vector.
And the PCR is coming in fact a good amplification...but around the single band there is a little glowing...what could be that.
apart from that can anyone tell why sometimes the band are not straight but slightly curved upwards from the ends. Even the marker ran along with it is curved. the gel was runned on 1% agarose.
beulah
And the PCR is coming in fact a good amplification...but around the single band there is a little glowing...what could be that.
apart from that can anyone tell why sometimes the band are not straight but slightly curved upwards from the ends. Even the marker ran along with it is curved. the gel was runned on 1% agarose.
beulah
Hi!! perhaps there is too concentrate. Try to put less material. Or improve the pcr. Perhaps the cube of electrophoresis. or if it is near a corner.
-capu-
Possible you have too much template. When bands curved in an electrophoresis the principal cause is to high voltage.
-merlav-
QUOTE (merlav @ Oct 14 2008, 05:21 AM)
Possible you have too much template. When bands curved in an electrophoresis the principal cause is to high voltage.
High voltage for the curved band!
High voltage causes gel to heat up. Uneven heating on the band will coz their movement to be slightly different abit. from edge to center.
therefore the dna will bend.
normally for confirmation of PCR success or restriction digest mapping checking it's ok. but for gel extraction maybe go lower the volt.
curved band not "FUN " to cut out
-Hanming86-