double sized PCR product from cloned cDNA - (Apr/26/2005 )
After I pcr the cDNA from my plasmid ,the product is double size of my cDNA.what kind problem could it be? Any suggestion will be appreciated!
-smzhu-
however unlikely, it could well be a concatemer of your cDNA, two copies ligated end to end and then into your vector.
-methylnick-
QUOTE (methylnick @ Apr 26 2005, 07:41 PM)
however unlikely, it could well be a concatemer of your cDNA, two copies ligated end to end and then into your vector.
Are you sure your construct is the correct size and the insert is inserted correctly? I would do a quick restriction digest.
-MaximinaNYC-
Dear friend,
Cut your band out and send for sequencing.
-yongyk-
QUOTE (MaximinaNYC @ Apr 26 2005, 06:37 PM)
QUOTE (methylnick @ Apr 26 2005, 07:41 PM)
however unlikely, it could well be a concatemer of your cDNA, two copies ligated end to end and then into your vector.
Are you sure your construct is the correct size and the insert is inserted correctly? I would do a quick restriction digest.
Thank you for your suggestions!I have done the restriction digestion and the result shows that the insert is a correct size.
-smzhu-
How big is the size of your PCR product? Is the size similar to the vector size? If yes, as discussed before, it could be the template.
-pcrman-
QUOTE (pcrman @ Apr 27 2005, 10:57 AM)
How big is the size of your PCR product? Is the size similar to the vector size? If yes, as discussed before, it could be the template.
The size is similar to the templet.So how can I get the product of my gene?Thank you very much for your good suggestion!
-smzhu-