Luckau Protocols:NanoDrop

Purpose
The NanoDrop is used to quantify genetic material. Although there are many different methods to quantifying DNA, the NanoDrop is a spectrophotometer that calculates absorbance of a sample across different wavelengths.

Protocol

 * 1) Assemble cafeteria tray
 * 2) * Kim Wipes
 * 3) * 2µL pipette
 * 4) * pipette tips
 * 5) * gloves
 * 6) * USB drive
 * 7) * water and buffer tubes
 * 8) * samples
 * 9) The NanoDrop is located in North Life Sciences, room 325A
 * 10) *[[Image:NanoDrop1000.jpg]]
 * 11) Open the NanoDrop software, "ND1000" on the desktop
 * 12) Choose "Nucleic Acids"
 * 13) Initialize the instrument
 * 14) * ensure upper and lower pedestal surfaces are clean by wiping with Kim Wipe
 * 15) * Place 2µL of NanoPure water on the lower pedestal
 * 16) * Lower the sampling arm and press OK
 * 17) * when it's done, wipe upper and lower pedestals with Kim Wipe
 * 18) Calibrate the instrument
 * 19) * Place 2µL of elution buffer on the pedestal
 * 20) * For Clark's lab, elution buffer is 10mM Tris-Cl, pH 8.4
 * 21) * Click "Blank"
 * 22) * when it's done, wipe upper and lower pedestals with Kim Wipe
 * 23) Measure sample
 * 24) * Place 2µL of sample on the pedestal
 * 25) * Enter sample ID
 * 26) * Click "Measure"
 * 27) * wipe upper and lower pedestals with Kim Wipe after each sample
 * 28) * Re-calibrate the instrument each 10 samples, or so, by going back to step 6 and clicking "Re-Blank" instead of "Blank"
 * 29) Save data
 * 30) * Click "Show Report"
 * 31) * Click "Reports", "Save Report"
 * 32) * Click "Export Report Table Only"
 * 33) * save to your USB drive, as "YYYYMMDD_NanoDrop", use "A" "B" "C", etc to designate multiple saved files in the same day
 * 34) Re-initialize the instrument each 30-50 samples, by going back to step 3
 * 35) * remember to save your data before your re-initialize
 * 36) Clean up thoroughly! Other people use this instrument!
 * 37) don't forget to enter data into spreadsheet

FYI

 * 260/280 = ratio of sample absorbance at 260 and 280nm, used to assess the purity of DNA and RNA; 1.8 is "pure" for DNA, <1.8 may indicate presence of contaminants
 * 260/230 = ratio of sample absorbance at 260 and 230nm, used as a secondary measure of nucleic acid purity; 1.8-2.2 is "pure", <1.8 may indicate presence of contaminants
 * NanoDrop User's Manual [[Media:NanoDrop1000-v3.7-UsersManual.pdf]]