Labeled cDNA Calculator

Kit Type Used:
Indirect Labeling
Direct Labeling, membrane based cleanup
Direct Labeling, resin based cleanup
 Type of RNA Used:
Total RNA
mRNA		   Cover Slip Size:
22 X 22 mm
24 X 40 mm
24 X 60 mm
 Spectrophotometer Path Length:
1 cm (10 mm)
1 mm
Volume of Purified
Labeled cDNA (µl)		 A260 A550 A650 Labeled cDNA Yield (ng) CyDye™ Incorporated (pmol) FOI Labeled cDNA Concentration (pmol/µl) Volume of Labeled cDNA to use for a Single Slide
Cy®3            
Cy®5            


 


For more details about microarrays, please see Troubleshooting Guide for Corning® UltraGAPS™ Slides and Corning® Pronto!™ Microarray Reagent Systems.


Notes:
Yield of cDNA (ng) = A260 X 37 ng/µl (1 A260 unit of ssDNA = 37 µg/ml) X volume (µl)
pmol dye incorporated: Cy®3 = A550 X volume (µl) / 0.15 (Cy®3 extinction coefficient at OD550 = 150,000 /M cm)
                                            Cy®5 = A650 X volume (µl) / 0.25 (Cy®5 extinction coefficient at OD550 = 250,000 /M cm)
FOI = pmol dye incorporated X 324.5 (average molecular weight of one kb of DNA in g/mol)/ ng cDNA

Standard spectrophotometers have a 1 cm (10 mm) path length. NanoDrop spectrophotometers have both 0.2 and 1 mm actual pathlength, but can report absorbance as 10 mm, 1 mm, or 0.2 mm pathlengths depending on the module being used.