Oligo Reconstitution and Use
All of our oligos are supplied lyophilized. These are stable at room temperature for extended period of time. We recommend the following reconstitution protocol. The solvent may be either sterile TE or sterile water depending on the established laboratory practice. After reconstitution store the stock solution at –80oC or –20oC.
The protocol given below is only to be used as a guideline and should not be substituted for any other specific protocol.
| Reconstitution |
Examples of Use |
Stock solution of 500 pmols/µl
[ 500 µM (micromolar) ] |
Polymerase Chain Reaction (PCR) |
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BSI provides the exact amount of nmoles of each oligo supplied on the tube and on the Oligo Report. Multiply the 'nmol' amount by 2 to arrive at the volume of solvent to be added.
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The final concentration of primers in a PCR reaction is usually 0.5 to 1 µM (micromolar).
This is equivalent to 0.5 to 1 pmol/µl. For a 100 µl reaction you would add 50 to 100 pmols. At BSI we use 0.5 pmol/µl; [0.5 µM (micromolar)].
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Example: 50.10 nmols × 2 = 100.2 µl
Dissolve the oligo in 100.2 µl to get 500 pmols/µl stock solution. Use as required.
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Stock solution of 100 pmols/µl
[ 100 µM (micromolar) ]
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Sequencing
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BSI provides the exact amount of nmoles of each oligo supplied on the tube and on the Oligo Report. Multiply the 'nmol' amount by 10 to arrive at the volume of solvent to be added.
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The final concentration of primer in automated sequencing is from 4 to 10 pmols (~0.05 - 0.1µg). Use the above dilution protocol to prepare a 100 pmols/µl [ 100 µM (micromolar) ] solution and then dilute 10 fold to get 10 pmol/µl solution. Use 1µl. (10 pmols)
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Example: 50.10 nmols × 10= 501 µl
Dissolve the oligo in 501 µl to get 100 pmols/µl stock solution. Use as required.
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Quick Conversion Table
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1 µM (µMolar)=1 pmol/µl (pico moles/µl). Example: 25 µMolar primer solution is 25 pmol/µl
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1 mM (mill Molar)=1 nmols/µl (nano moles/µl)
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The molecular weight of the oligo = Length of oligo × 330 dalton
For an 20 mer oligo, the MW = 30 × 330 = 6600
Example: 6600 g = 1 mol, 6.600 µg = 1 nmol
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