in this video we will show you how to operate the Nanodrop Spectrophotometer
specifically demonstrating how this equipment can be used to measure concentration of nucleic acid samples
the Nanodrop Spectrophotometer can be used to directly measure the concentration and purity of protein, DNA or RNA in the sample
When working with DNA or RNA it's important to always keep the samples on ice and wear gloves to prevent sample degradation
to open the nanodrop software click on the ND1000 icon
Today, we'll be working with Nucleic acid so let's select the nucleic acid option
The Nanodrop should be gently wiped down with the Kimwipe and some dd water. Now we're ready to use the Nanodrop
first using a pipetman p2 or p 20, pipette 1 microliter of RNase free water onto the sensor pedestal of the nanodrop fully covering it without generating any bubbles
Please note that any volume between 1 microliter to 2 microliters is acceptable
Then gently pull down the Nanodrop arm onto the drop of liquid
Select ok on the computer and this will initialize the equipment
Next we can take the blank reading
to blank the Nanodrop pipette one to two microliters of the solution used to elute your DNA or RNA sample onto the sensor pedestal and gently lower the Nanodrop arm
then click blank to read
When the reading is complete lift up the Nanodrop arm and clean the sensor pedestal with the Kimwipe
Please note the Nanodrop requires re-blanking after every 10 sample readings
Now we're ready to read the sample
pipette 1 to 2 microliters of your DNA or RNA sample onto the sensor pedestal and gently lower the Nanodrop arm
Go to the drop down menu and Select either DNA or RNA depending on your sample
in this example we're using DNA. Then click measure to read
When finished clean the sensor pedestal gently with Kimwipes, repeat this step for additional samples
Name your sample by typing in the sample ID box. Click show report and export to excel or copy to your lab notebook
Here is an example of a DNA reading
the ratios 260 over 280 and 260 over 230 are especially important
The 260 over 280 Ratio is used to determine protein contamination of a nucleic acid sample
the 260 over 230 Ratio indicates presence of organic contaminants
for pure DNA and RNA the 260 over 280 ratio should be within the range of 1.8 to 2.1
the closer it is to 2.1 the more ideal because this indicates that the sample is not protein contaminated
As for the 260 over 230 ratio any value lower than 1.8 will indicate significant contamination
while pure samples should have a value close to 2.0
As mentioned the Nanodrop Spectrophotometer can also be used for protein concentration measurement
an example can be found in the case in a solution video which will be linked above in this video and in the description box below
When you're finished with everything clean the Nanodrop with a kimwipe and some dd water
Store the Nanodrop by inserting a piece of folded Kimwipe between the arm and sensor pedestal
Thank you for watching