Nanodrop(Protocol( - Weber Labweberlab.byu.edu/Portals/105/Protocols/Nanodrop...
Transcript of Nanodrop(Protocol( - Weber Labweberlab.byu.edu/Portals/105/Protocols/Nanodrop...
Nanodrop Protocol 9/25/2013 TO
Purpose The main reason to run a nanodrop is to find the concentration of your DNA (or protein). We run a nanodrop after each miniprep cleanup we do and before we run any experiment where we need to know the amount of DNA we have (such as restriction digests, ligations, etc.). Protocol
1.) Prep your DNA you wan to nanodrop (miniprep, PCR cleanup, gel extraction, etc.).
2.) Assemble all tools you need: a. 2.5 pipette b. tips c. DNA sample d. Blank of some sort. For example, if you eluted DNA, then use the
elution buffer as your blank. e. Marker
3.) The nanodrop machine is in the Grose lab.
i. 4.) Turn on ND-‐1000
i. 5.) Load 2μL of ddH2O to wash.
ND-‐1000
ND-‐1000
i. 6.) Exit out, blot with a kimwipe, and reload another 2μL of ddH2O
i. 7.) Blot and Load 2μL of the blank 8.) Now you are ready to measure. To make sure you are correctly calibrated,
load another 2μL of your blank and hit “Measure.” This measurement should not be off by more than 1 ng/μL.
i. 9.) Label your sample under “sample ID”. Blot, load 2μL of your sample, and hit
measure. Save a copy of your graph after each sample under the “Weber” folder on the desk top.
Notes Your 280/260 measurement should be about 1.80. Samples should read about 100-‐300 ng/μL Your graph should look like the graph to the right.
Load Samples Here
Blot to Clean
Measure, Blank
Nanogram r
eading