AlanFung:Protocol/OliGreen ssDNA Stain: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 No edit summary |
>Alan6017518 |
||
Line 22: | Line 22: | ||
==Step 2: Oligonucleotide Standard Curve== | ==Step 2: Oligonucleotide Standard Curve== | ||
*Dilute the oligonucleotide standard, provided at 100ug/mL to 50 fold in TE to make the 2ug/mL working solution | |||
*For low-range standard curve from 100pg/mL to 50ng/mL prepare a 20-fold dilution of the 2ug/mL oligonucleotide solution to yield a 100ng/mL oligonucleotide stock solution | |||
*Dilute 100ng/mL oligonucleotude stock solution |
Revision as of 20:52, 28 May 2009
Probe Preparation
Samples & Materials
- Quant-iT OliGreen ss DNA Assay Kit (Cat. no. O11492)
*Quant-iT OliGreen ss DNA Reagent (Component A), solution indimethylsulfoxide (DMSO) *20X TE Buffer (Component B) *Oligonucleotide Standard (Component C) 18-base M13 sequence primer, with the sequence 5'-TGTAAAACGACGGCCAGT-3'
- Sterile, distilled, DNase-free water
- 96 well microplates
- Microplate Reader
Overview
Procedures
Step 1: Preparing Reagent
- Allow the reagent to warm to RT before opening the vial
- Prepare an aqueous working solution of the Quant-iT OliGreen reagent by making a 200 fold dilution of the concentrated DMSO solution in 10nM Tris-HCL
1mM EDTA pH 7.5 (TE)
Step 2: Oligonucleotide Standard Curve
- Dilute the oligonucleotide standard, provided at 100ug/mL to 50 fold in TE to make the 2ug/mL working solution
- For low-range standard curve from 100pg/mL to 50ng/mL prepare a 20-fold dilution of the 2ug/mL oligonucleotide solution to yield a 100ng/mL oligonucleotide stock solution
- Dilute 100ng/mL oligonucleotude stock solution