TITLE: Nanodrop Lite (Shared Equipment Lab)
AUTHORS: Nimalka M Weerasuriya

[DESCRIPTION]
How to use the Nanodrop Lite.

[BEFORE_START]
It is best to use a precision pipettor (0-2 μl) with precision tips to ensure that sufficient sample (1-2 μl) is delivered. Lower precision pipettors (0-10 μl and larger) are not as good at delivering 1 μl volumes to the lower pedestal. If the user is unsure about the sample characteristics or pipettor accuracy, a 2 μl sample volume is recommended.



[GUIDELINES]
A 1-2 μl sample is pipetted directly onto the lower pedestal. A fiber optic cable (the source fiber) is embedded within the lower pedestal while a receiving fiber is imbedded in the upper pedestal and the arm. Light emitted from the LEDs traverses the sample and absorbance is measured using a silicon photodiode.

[STEPS]
SECTION: Setup
1. Open arm or push any button to wake instrument.
Using the bottle of deionized water, slightly wet a Kimwipe and clean the lower and upper pedestal (metal prongs).

 


SECTION: Nucleic Acid Measurements
4. Select the appropriate application from the Home screen (DNA or RNA). For DNA measurements, select either the dsDNA or ssDNA assay.
SECTION: Nucleic Acid Measurements
5. Following the on-screen instructions, establish a blank by pipetting 2 µL of the blanking buffer (RO or elution solution your DNA is in) onto the bottom pedestal, lower arm and press Blank.
 


SECTION: Nucleic Acid Measurements
6.1. When measurement is complete, raise the arm and wipe the buffer from both the upper and lower pedestals using a dry laboratory wipe.
SECTION: Nucleic Acid Measurements
8. Measure sample by pipetting 2 µL of sample onto the bottom pedestal, lower arm and press Measure.

 

 
SECTION: Nucleic Acid Calculations
9. Nucleic acid concentration ranges:
    ABC1Sample Type LowerLower Detection LimitUpper Detection Limit2dsDNA4.0 ng/ul1500 ng/ul3ssDNA2.6 ng/ul990 ng/ul4RNA3.2 ng/ul1200 ng/ul

 

SECTION: Setup
2. Select measurement type:
SECTION: Protein Measurements
3. Select Protein from the Home screen.
SECTION: Protein Measurements
4. Select the appropriate assay type: 1A/cm=1mg/ml, IgG, or BSA.
SECTION: Protein Measurements
5. Following the on-screen instructions, establish a blank by pipetting 2 µL of the blanking buffer onto the bottom pedestal, lower arm and press Blank.
 


SECTION: Protein Measurements
6. When measurement is complete, raise the arm and wipe the buffer from both the upper and lower pedestals using a dry laboratory wipe.
SECTION: Protein Measurements
7. Confirm Blank by pipetting a fresh aliquot of blanking buffer onto the bottom pedestal, lower the arm and press Blank.
SECTION: Protein Measurements
8. When measurement is complete, raise the arm and wipe the buffer both the upper and lower pedestals using a dry laboratory wipe.
 
SECTION: Protein Measurements
9. Measure sample by pipetting 2 µL of sample onto the bottom pedestal, lower arm and press Measure.

SECTION: Protein Measurements
10. Wipe the upper and lower pedestals using a dry laboratory wipe and the instrument is ready to measure the next sample.
SECTION: Protein Calculations
11. Protein concentration detection limits
 
 
 

SECTION: Nucleic Acid Measurements
7. Wipe the upper and lower pedestals using a dry laboratory wipe and the instrument is ready to measure the next sample.

SECTION: Nucleic Acid Measurements
3. Heat high molecular weight DNA samples to 55 °C and gently vortex before measurement. Due to the small volumes required by the NanoDrop Lite, it is important to ensure that the sample being measured is homogeneous. Field experience has shown that samples containing large molecules such as genomic or lambda DNA are particularly susceptible to heterogeneity.
SECTION: Nucleic Acid Measurements
8.1. The resultant A260/A280 ratio for the nucleic acid being studied will be approximately equal
to the weighted average of the A260/A280 ratios for the four nucleotides present. 
It is important to note that the generally accepted ratios of 1.8 and 2.0 for DNA and RNA are
“rules of thumb.” The actual ratio will depend on the composition of the nucleic acid.
 

SECTION: Nucleic Acid Measurements
5.1. When measurement is complete, raise the arm and wipe the buffer from both the upper and lower pedestals using a dry laboratory wipe.
SECTION: Nucleic Acid Measurements
6. Confirm Blank by pipetting a fresh aliquot of blanking buffer onto the bottom pedestal,
lower the arm and press Blank.