We are planning to buy Topo TA cloning kit without the competent cells due to higher cost of the kit. We have home made XL1 blue E.coli chemical competent cells in our lab. We can also prepar some DH5alpha cells, if need be. My doubt is whether or not the TopoTA cloning in compatible with XL1 Blue E.coli competent cells that we made in our lab? Is there any special reason of using Top 10 cells with TopoTA cloning kit provided by the company? Can we use TopoTA cloning kit and transform into XL1 blue cells or DH5alpha cells?
Dear Saheli
in principle you can certanilly use DH5aplha and XL1 bue for Topo cloning.
I used several time DH5aplha and it work well.
However i suggest to you to evaluate an enzime free cloning approach as PIPE (Polimerase incomplete primer extention) cloning.
Before know about PIPE i was also a fun of Topo cloning and Gateway but once in discover PIPE my life changed a lot. PIPE allow to you to use any vector with out any restrinction and it is very simple and fast.
you can find more details about it on my blog:
ProteoCool https://proteocool.blogspot.com/
ProteoCool n°1 (Cloning methods overwiev) and
ProteoCool n°3 (Primer Design: PIPE Cloning)
good luck
Manuele
Hello Saheli,
I used TOPO TA with DH5alpha, TOP10, and TG2 cells and worked well with these E. coli competent cells. I prefer TG2 or DH5alpha cells because the size of the colonies is bigger than the size of TOP10 colonies.
So, in principle, you can try the XL1 Blue cells with TOPO TA. It should work.
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