DNA Quantification - Oly Oyster gDNA-01 for RAD Sequencing (from 20141014)

Quantified the gDNA isolated 20141014 using the Quant-iT dsDNA Broad Range kit (Life Technologies).

Used half the recommended volume of buffer (100uL instead of 200uL), as well as half the recommended volume of dye (5uL instead of 10uL). Prepared a master mix of the buffer/dye solution, containing a 1:200 dilution of the dye:

Buffer: 14745uL

Dye: 75

100uL of the solution was added to the appropriate wells in two 96 well, black plates with a multichannel pipette.

5uL of each standard was added to the designated wells (see the plate layout in the Results below). Standards were run in duplicate.

5uL of each samples was added to the designated wells.

Samples were quantified using a plate reader.


Raw fluorescence was collected and an equation for the standard curve was determined for each plate. Concentrations were calculated from that equation (see linked spreadsheet below).

Concentrations of each sample in their corresponding locations in the Oly Oyster gDNA-01 plate are here: 20141022-OlyRADdnaConcentrations

Additionally, the volumes needed for 500ng of each sample the required starting amount for the RAD sequencing protocol) are also on the spreadsheet linked above.