I am trying to clone h 533 bp fragment in a 21000bp vector. What should be appropriate ratio of vector to insert to use for successful cloning.
A good place to start is a 3x molar amount of insert to vector.
So if you use INS(ng) =Vector ( ng) x (INS size/Vector size) all x3
so in your example the amount of insert would be given for X ng of vector would be
INS (ng) = Xng x( 533/21000) x 3
Small fragments are easier to clone than long ones.
I believe this is due to the higher diffusion rates of small fragments, which results in more frequent productive collisions between vector and insert.
In your case, I'd be inclined to use a rather low insert:vector ratio (and don't forget to divide DNA concentration by length), otherwise you may end up with a lot of double inserts. But if the percentage of recombinants is less than about 30% you are probably safe.
Ritika Vishnoi if you read carefully the answer that Paul Rutland provided is to calculate based on molar ratios. This will be independent of size but of course you need to take into account size and amount in order to calculate it.
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