Dinh/NOTES/2009-12-4

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Quant-iT(TM) ssDNA Assay Kit

Materials

  1. Quan-iT ssDNA Assay Kit (cat. # Q32851, containing:
*Quant-iT ssDNA reagent, solution in DMSO
*20xTE (200 mM Tris HCl, 20mM EDTA, pH7.5)
*Oligonucleotide standard (100ug/ml = 100ng/ul)
  1. Quan-iT OliGreen Assay tubes (500ul)
  2. Sterile, distilled, DNase free water

Table 1. High range standard curve

Volume (ul) 1xTE Volume (ul) of 2ug/ml Oligomer stock Volume (ul) of diluted Working Solution Final Oligomer Conc.
Standard #1 0 250 250 1ug/ml
Standard #2 125 125 250 500ng/ml
Standard #3 225 25 250 100ng/ml
Standard #4 247.5 2.5 250 10ng/ml
Standard #5 250 0 250 blank

Protocol

1. Preparations

a. Prepare 1xTE buffer (1 ml per each sample to be assayed)
b. Prepare working solution by 200 fold dilution of OliGreen ssDNA reagent (in DMSO). 
For best results, use working solution within few hours of preparation.
c. Dilute oligonucleotide standard 50 fold to 2ug/ml