concentration of vector in ligation reaction - (Jul/29/2007 )

The vector which is used in ligation reaction should have a high concentration?? Is it true? If my vector is onle 5.5 microgram/ microliter, it still can be used in ligation reaction?

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If the vector is really 5.5 micrograms, its a lot. You need to dilute it to use for ligation. For a normal ligation, I try to keep my total DNA (vector plus insert) around 30 ngs. For some I would go upto 200ng but this is rare.

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use 1:3 or 1: 5 ration of vector to insert. If blunt end ligation you use 1:10 ratio. You can still use your vector but you have to dillute your vector way down to 50ng/uL

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sorry....typing mistake: the concentration should be 5.5 nanogram/microliter

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Well, Normally I would use 20-30 ng of vector. Here, I would simply use 4 ul of the vector for ligation.

Good Luck !!!

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