Help with cloning

lucia akinyi akinyistar at googlemail.com
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,
Lucia.

-- 
The good Gracious Lord never forsakes His people Amen.

Lucia Akinyi Odhiambo


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