PCR cloning kits allow to clone a PCR product in a variety of vectors through a simple blunt-ended ligation.
However, would this still work with PCR products containing modified ends (obtained with modified oligos such as biotinylated of fluorescin-labeled primers) ? I only need the DNA sequence to be inserted into the vector, not the label.
Thanks
Why shoud anyone take a chance with an expensive biotinylated oligo when native oligos ar dirt cheap and can be obtained from commercial companies in a matter of days ?
lots of types of biotins linked oligos, not sure whether yours affect the ligation.
you could try it.
In order for the ligase to form a phosphodiester bond, the 5-hydroxyl of the oligonucleotide mast contains a phosphate group. In your case when biotin is linked to 5-hydroxyl of oligonucleotide, I blut-end cloning is not an option.
Tried it and it worked perfectly fine with 2 different modifications (biotinilated oligos and fluorescin-tagged oligos) ! I have the same amount of transformants as another blunt cloning with regular (non-modified) PCR products.
Dear Martin
Which type of cloning kit do you used ? Ligase or topoisomerase based?
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