Protocol Online logo
Top : Forum Archives: : Molecular Biology

Molar Ratio of Inser to Vector - (May/29/2007 )

H, Im trying to ligate an insert of just 46bp to a BamHI/HindIII cut pUC19. I wonder if its possible to use molar ratio calculation of such a small insert. Usually , the inserts are around few hundreds bp to >1kb. Using nano drop ive determined my vector to be only 9ng/ul after i gel purified my double digested pUC19. While the insert is around 900ng/ul according to nanodrop.

-loserDNA-

you could use the molar ratio calculation for such a small insert. I have used it to clone a linker into pBS.

-scolix-

i agree with scolix.
Go for 20ng of plasmid and 2-3┬Ál of your PHOSPHORYLATED insert.
normally, should be not that difficult to get good ones

-fred_33-

http://www.promega.com/biomath/calc01.htm

you can use this calculator

-T. reesei-