Spectrophotometer, Nanodrop PCR

NANODROP PCR (B412)

Rules and Guidelines

BEFORE USE:

1. New user MUST be trained by the captain or present users

QUICK START:

1. Double click on the desktop NanoDrop™ 2000 software icon and select the application of interest. Follow the prompts for instrument initialization.

2. Ensure Add to report is selected in the left pane to automatically include all measurements in the saved report.

3. Establish a Blank using the appropriate buffer. Pipette 1-2 ul of the blanking buffer onto the bottom pedestal, lower the arm and click the

Blank button. The blank solution is generally the same buffer that the molecule of interest is suspended or dissolved in.

− For the NanoDrop 2000c model, select the Use cuvette box to make measurements with a cuvette.

− Insert the cuvette noting the direction of the light path indicated by the etched arrow.

− The arm must be down for all measurements-including measurements made with cuvettes.

− The optical path is directed 8.5 mm above the bottom of the cuvette.

Refer to the manufacturer for volume recommendations.

4. Wipe away the blank and enter the sample ID in the appropriate field. Pipette 1-2 ul of sample and hit Measure.

− It is recommended that a fresh aliquot of sample be used for each measurement.

After a measurement:

− Wipe both measurement pedestals using a dry, lint-free laboratory wipe and the instrument is ready for the next sample.

− When using the cuvette option, remove the cuvette, rinse thoroughly and dry between samples.

Blanking Cycle

It is generally recommended that an aliquot of the blanking buffer be measured as if it were a sample. This will confirm that the instrument

is working well and that any sample dried down from previous measurements is not a concern. To run a blanking cycle,

perform the following:

1. Load an aliquot of the blank onto the lower measurement pedestal and lower the sampling arm into the down position.

2. Click on the Blank button to store the blank reference.

3. Analyze a fresh replicate of the blank as though it were a sample by selecting Measure. The result should be a spectrum that

varies no more than 0.04 A (10 mm absorbance equivalent).

4. Wipe the blank from both measurement pedestal surfaces and repeat the process until the spectrum is within 0.04 A (10 mm

path).

Although it is not necessary to blank between each sample, it is recommended that a new blank be taken every 30 minutes when

measuring many samples.

REFERENCE MANUALS:

--Jessica Metzger 18:09, 17 December 2012 (EST)