Dinh/NOTES/2009-12-4
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Quant-iT(TM) ssDNA Assay Kit
Materials
- 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)
- Quan-iT OliGreen Assay tubes (500ul)
- 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