Download APC004 DNA Quantification/Nanodrop

STANDARD OPERATING PROCEDURE: #APC004, Rev 2
Title: Quantification of Genomic DNA using the Nanodrop-1000 UV-Vis spectrophotometer.
Effective Date: 18 Sept 2013
Approvals (Signature and Date):
Selena Healy
Sally Cudmore
Author – (Signature)
1.0
Molecular Biology grade dH2O (Sterile) (e.g. Sigma)
DNA Rehydration Solution (e.g. Qiagen/Promega)
Lens Cleaning Tissue
P10/P2 Pipettors and suitable tips. (Gilson). (Lab 5.27 BSI)
Nanodrop-1000 UV-Vis spectrophotometer
HEALTH AND SAFETY CONSIDERATIONS
5.1
6.0
It is the responsibility of all users to receive training on the equipment, and to keep records to
demonstrate proof of training received.
MATERIALS AND EQUIPMENT
4.1
5.0
The scope of this SOP is to provide comprehensive detail to the quantification of genomic DNA using
the Nanodrop UV/Vis Spectrophotometer.
RESPONSIBILITY
3.1
4.0
The purpose of the SOP is to Quantify Genomic DNA..
SCOPE
2.1
3.0
Date: 20 Sept 2013
PURPOSE
1.1
2.0
Technical Authority (Signature)
Glasses and labcoats are required during use of this equipment.
DOCUMENTATION REQUIREMENTS
6.1
Please refer to the manual of the spectrophotometer prior to and during use.
7.0
PROCEDURE
7.1
Clean the Nanodrop Instrument sampling pedestal with a clean piece of lens tissue with dH2O.
Dispose of the tissue in the normal bin, it is only paper!!
Note: Ensure that only lens tissue is used. Ordinary tissue can damage the Nanodrop.
Cleaning
7.2
Start-up the Nanodrop application (3.01) on computer in Lab 5.27., and select Nucleic acid.
7.3 The screen below will appear, follow the instructions and then add 1l of dH2O on to the Nanodrop sampling
pedestal, see images below.
(Wear Gloves, unlike above!)
7.4 Select Record and then Blank from the buttons and blank the instrument with your solution used for
resuspending your DNA samples, a sample ID can also be added but you will have to edit it for each sample read.
7.5
7.6
A reading of 0.0 ng/l should appear in the concentration box.
Add Your DNA sample to the Nanodrop and click Measure. A measurement will appear. If the sample is
very high in concentration it is advisable to dilute it 1:5 or 1:10 as Genomic DNA can be very viscous and
may yield in incorrect readings.
7.7 Continue adding DNA samples Wiping the pedestal clean before each new sample, you can Blank or re-read
samples if required, just ensure to change the sample ID each time.
7.8 When you are finished click Exit (top right button)
7.9 A Nanodrop Nucleic Acid Report will appear, Save the report by clicking on the button, it is advisable to add
your initials and date so as it can be found. The data can be exported to Microsoft Excel relatively easily.
Note - RNA measurements are similar, but remember to check your ratio of 260/280 and also use the Agilent
Bioanlayser for integrity of assessment, this is most important.
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