after we have done transformation ,is it possible to compare colonies on plates, i mean is it possible to find that fragment-vector ligated are bigger than self-vector ligated?
you can do this in three way I will put them from the fastest to slowest:
1- if you are using a vector that supports blue-white screening you can use xgal+IPTG to screen for vector ligated colonies (the colonies with insert will be white while self ligated vector will be blue) followed by restriction digest to confirm.
2-Colony PCR (one primer in the backbone and one in the insert) followed by restriction digestion to confirm.
3- Restriction digestion after miniprep, however, I would not recommend this since depending of how your ligated your insert (for example single cut vectors) you might get a high proportion of self ligated vector.
Dear Abire
you can simply perform a colony pcr by:
- using both primers which anneal on the plasmid backborne and distunguish self ligate plasmid from the insert containing plasmid from the size of pcr band;
- using a primer annealing on plasmid and the other on the insert.
in this way the positives bands correspond to the right clones.
you can find a detailed protocol on my blog : https://proteocool.blogspot.com/
ProteoCool n°4 at page1
good luck
ciao
Manuele
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