According to manual the formula is
ng = ( x fmol )( N )(660 fg/fmol)(1 ng/106fg)
where N is the size of the DNA in base
pairs, and x is the number of fmoles. so , if my entry clone is 2817bp and insert size is 1374bp ,then N=4191bp and x=10. that means i will need 28ng/microliter DNA. Am I right??
Another question, According to manual I have to use 20fmol destination vector and the volume is 1 microliter. Can I use more than 1 microliter DNA say- 2 microliter containing 20fmol DNA? Because my Destination vector DNA is low concentrated.
Hi there,
Your formula, although a bit complicated, looks OK. More generally, mass of DNA in g=number of moles*number of bp*660g. 660 being the average MW of a bp. The rest is about conversion.
Provided the final volume is conserved there is no problem as what actually matters is the final concentration of reagents in the mixture.
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