Question

What is considered a good nanodrop value (nucleic acid concentration) in ng/μL?

Answer 1

This depends entirely on what you are doing. If you extracting DNA/RNA, then it depends on whether you are extracting from blood or tissue. If you are doing a specific procedure, then see the protocol for that procedure. I know many cloning procedures, for example, only require about 1-50ng/uL, and in some cases even less. Again, this depends on what you are doing.

Answer 2

Sorry, I meant to clarify, but I got distracted. I took the genetic material from bacterial clones, which (hopefully) incorporated the genes from a PCR we ran a couple weeks ago. I mini-prepped the bacteria today, and I will use the resulting genetic material to make a standard curve, which will later be used to measure gene expression between tissues. I would ask the PI/head of the project about how much we need, but she's out of town for the next few days and I'm on my own. Does the ideal value still depend on other factors, or is there a standard density to shoot for?

Answer 3

Ok, are you using Datsanko-Warner protocol? PCR product based cloning is a pretty broad base, but i have done some similar things...

Are you looking for the concentration of PCR product to add to your cultures or is this a plasmid concentration? If you are doing any ligation then this also gets a bit tricky.

If you are looking for how much to clone into your competent cells, then it should be in the protocol which you can find online. If this is the case, just google your competent strain (ex: DH5 alpha or JM109) to find the recommended concentration of PCR product to add. If you are using electroporation or sonoporation to clone this into cells that have not been made competent, concentrations again vary. Otherwise, get back to me and i will try to help!

sorry for all the questions, dont want to give bad advice!