Help with cloning

lucia akinyi akinyistar at
Thu Jun 29 02:36:41 EST 2006

I want to clone a 500bp insert into pGEMT vector. After overnight ligation
and transformation of the competent cells and growing on LB agar ampicillin
plates overnight, i get colonies which when i screen i get by pcr
amplification i get a 500bp band. I then grow the same colonies (which gave
the 500bp band) in LB broth medium overnight and after purification of the
plasmid using the QIAGEN kit, i run a pcr again but this time a get a band
approximately 2500bp. I am sure it is not the vector because the vector is
approximately 3000bp. Does anyone have an explanation for this disparity?

Thank You,

The good Gracious Lord never forsakes His people Amen.

Lucia Akinyi Odhiambo

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