Smearing on ategarose gel of real time pcr product??? - (Jun/14/2010 )
HERE ITS DR DANZ....
I M WORKING ON EXPRESSION STUDY OF TLR GENE IN MASTITIS USING REAL TIME PCR
I AM WORKING ON AB7500 FAST REAL TIME SYSTEM AND GETIING GOOD EXPRESSION-
GOOD AMPLIFICATION PLOTS AND MELT CURVE.
THE SAME PRODUCT WHEN DONE BY NORMAL PCR SHOWS BANDC OF APROPRIATE SIZE(200 BP)
I M USING QUIAGEN QUANTIFAST SYBR GREEN MASTER MIX
ANY SUGGESSION IS APRECIABLE
Could your qPCR buffer be affecting the gel? There may be something in the qPCR reaction mix that makes running the gel problematic. Check the protocol and instructions which came with your qPCR reaction, there might be some clues there. Perhaps you could try cleaning your PCR product? This might help?