I transformed a vector (pCMV201), used for virus-induced gene silencing (VIGS), into DH5-alpha E.coli strain with traditional heat-shock method. After recovery for 40 mins in total of 0.5 mL of liquid LB, I centrifuged them under 8K rpm for 5 mins. I resuspended the cell-pellet with 0.1 mL of liquid LB, serial diluted them for 10x and 100x, then plated 3 LB-plates with each dilution. However, after 16 hrs the following day, they were all smear growth with 1x been thickest, followed by 10x and then 100x. It's the second time I'm doing now and it's still showing the same. However, this time I don't wish to discard the plate immediately, is there any suggestion as to use the loop and plate them or do I just do a third transformation? I don't have glycerin stock since the lab who donated this pCMV201 only gave us in the form of purified plasmid. Thanks.

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