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# Equation for calculating insert:vector ratio in ligation reaction

5 replies to this topic

### #1 sagar

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Posted 26 February 2009 - 09:27 AM

hi all

i am very much in need for the equations to carry out calculations before performing ligation, can any one plz help?

### #2 T C

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Posted 26 February 2009 - 08:03 PM

Hey

Equations? calculations? for ligation?

I am not sure if I understand what you mean to convey.

TC

hi all

i am very much in need for the equations to carry out calculations before performing ligation, can any one plz help?

### #3 scolix

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Posted 28 February 2009 - 08:54 AM

Do you need the ligation recipe or how to calculate the vector-insert ratio? Please be more specific.

### #4 planktonica

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Posted 04 March 2009 - 01:16 PM

insert mass in ng = 6 X (insert lenght in bp/vector lenght in bp) X vector mass in ng

You may change the 6 above depending on the actual ratio you want to get (sometimes a 3:1 works best than 6:1)

### #5 Nrelo

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Posted 05 March 2009 - 12:38 AM

insert mass in ng = 6 X (insert lenght in bp/vector lenght in bp) X vector mass in ng

You may change the 6 above depending on the actual ratio you want to get (sometimes a 3:1 works best than 6:1)

One thing to mention, you have to get pure vector and insert before applying this equation. Sometimes RNA contamination will lead to incorrect calculation.

Try 3:1 and 1:1 insert to vector ratio, these two ratio usually rock in ligation reaction.

### #6 frg

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Posted 08 March 2009 - 09:10 PM

Hi Sagar,

Go to the 'Biomath Calculator' website using the following link.

http://www.promega.c...ath/default.htm

Click on Ligations: Molar Ratio of Insert to Vector

Enter the parameters of your ligation and it will calculate the relative amounts for you.

Easy!