DNA estimation - (Aug/21/2005 )
Hi, my questin is why we need to take reading of the DNA extraction by spectrophotometer or flurometer (DNA/RNA calculator). i know that for further work as PCR or RE, but why we need to take ratio, purity, protein concentration what is the benefit for these reading?
i am sorry. i don't know the answer to ur question as i am relatively new to this field too. however, i have a question on dna estimation. how to we estimate the concentration by comparing the bands of interest with the dna ladders bands. for example, i am using the promega 1kb dna ladder...and i use 5ul which has 500ng. there are 13 bands in the ladder...but some of them are of higher intensity than others...so how i find out the concentration of these ladder bands and then estimate my band of interest? pls help...
For some applications, for instance transfections, you need pure DNA without contamination of other things.
You can estimate the concentration of your DNA by comparing it to the marker, you should have an insert in the package (when you receive the marker) that states the concentrations for every band. Otherwise, you can look online, at the website of the company that has sold you the marker, it should have the information you're looking for.