Team:Aachen/Wetlab/Protocol9

Resuspension of IDT DNA Oligonucleotide

Procedure:


  1. Resuspend oligos in TE (Tris-EDTA) buffer (10 mM Tris; 0.1 mM EDTA; pH 8.0) as this buffer will maintain a constant pH. Alternatively, use nuclease-free water (Not DEPC)

  2. Resuspend dry oligonucleotides to a storage stock concentration of 100 uM and then dilute a portion of this to create working stock solutions.

storage stock concentration:

  1. Suspend the primers in the IDT tubes

  2. To make a 100 uM concentration stock solution (as backup in the IDT tube): Take the number of nmoles in the tube (from the tube-label) and multiply that by 10. This will be the number of μL buffer to add to get a 100 μM solution.

  3. Incubate for at least 15 minutes.

  4. The tube can be stored at -20 ℃.

PCR working stock:

  1. Take a new tube for this stock.

  2. To get 100 uL 5 uM solution, mix 5 uL of 100 uM oligo stock with 95 uL of water or TE buffer.

  3. Use the 5 uM stock for the PCR Mastermix and store the rest at -20 ℃.

Tips:

If you want a different concentration than 100 uM or 5 uM you can use the IDT Dilution Calculator (IDT Dilution Calculator)