1. You just received a dry oligonucleotide in a 2 mL tube from IDT. The oligo th
ID: 182353 • Letter: 1
Question
1. You just received a dry oligonucleotide in a 2 mL tube from IDT. The oligo that you ordered has the sequence TGCAGGACCAGAGAATTCGAATACA. The tube says “80.7 nmol”. How much TEz buffer should be added to make a 100 uM stock solution?
2. The oligo you received is the “ACA2” oligo that we use to amplify libraries. How much 10 uM PCR primer working solution can you make from the tube you just received?
3. IDT host a very useful “oligo-analyzer” tool (https://www.idtdna.com/calc/analyzer). Enter the sequence for ACA2. What is the melting temperature of ACA2 in a solution that contains 200 mM NaCl? What is the melting temperature in a solution that contains 20 mM NaCl and 10 mM MgCl2? Which is higher? What does “melting temperature” mean?
4. The analyzer spits out the molecular weight of the ACA2 oligo. How much mass, in units of milligrans, micrograms and nanograms of ACA2 oligo is in our 80.7 nmol?
Explanation / Answer
Answer:
Amount of oligonucleotide in the tub received from IDT = 80.7nmol
1) How much TEz buffer should be added to make a 100 uM stock solution?
2. The oligo you received is the “ACA2” oligo that we use to amplify libraries. How much 10 uM PCR primer working solution can you make from the tube you just received?
3. IDT host a very useful “oligo-analyzer” tool (https://www.idtdna.com/calc/analyzer). Enter the sequence for ACA2. What is the melting temperature of ACA2 in a solution that contains 200 mM NaCl? What is the melting temperature in a solution that contains 20 mM NaCl and 10 mM MgCl2? Which is higher? What does “melting temperature” mean?
4. The analyzer spits out the molecular weight of the ACA2 oligo. How much mass, in units of milligrams, micrograms and nanograms of ACA2 oligo is in our 80.7 nmol?
Molecular weight of the ACA2 oligo based on analyzer = 7708.1 g/mole
Amount of ACA2 oligo = 80.7 *10-9 mole
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