1) If there are no single stranded oligos in the -20 freezer they need to be ordered from IDT Technologies. See the page on doing this.
2) To make the adapters there are four oligonucleotides:
| Name | Sequence 5'-3' |
| Eco A1 | CTC GTA GAC TGC GTA CC |
| Eco A2 | AAT TGG TAC GCA GTC TAC |
| Mse A1 | GAC GAT GAG TCC TGA G |
| Mse A2 | TAC TCA GGA CTC AT |
*Note that these primers do not have a Phosphorylated tail*
3) If the oligos are new and dried, then you must dissolve them in 1X STE (10mM Tris pH 8.0; 50mM NaCl; 1mM EDTA) to a high concentration of 650 uM (uM is the same as pmoles/ ul.
1) Put equimolar amounts of each Eco oligo into one 0.5 ml tube, and each Mse oligo into another 0.5 ml tube.
2) Heat the tubes to 95C in a heat block for 3 min.
3) Remove the entire block with the tubes and put it on the bench top to cool to room temperature slowly.
4) Now you have a concentrated stock of annealed adapter at 325 uM. To make the working concentration dilute the Mse adapter to 50uM and the Eco adapter to 5uM.
4) Store at -20 C