Ordering barcoded primers
- Contact IDT or your primer provider to get a quote prior to ordering.
- Barcoded primers are ordered in 96-well plates at the 100-nmol scale.
- Make sure to select the option to receive the same amount per well of each primer (do not get full yield). This is generally the 8 nmol/well option.
- Select the rows option for load scheme: [Rows [A01,A02,A03]]
- If you will be archiving plates of primers for later use consider selecting the option for replicate plates.
Resuspension of primers
- Resuspension of primers is a critical step and must be done with ultra-pure water under sterile conditions. It is recommended that resuspension be done in a hood.
- Stock plates are generally 100 µM and are aliquoted and diluted to 10 µM for use.
- When making stock plates it is a good idea to make multiple replicate plates so that there are backup plates of primers.
- The primer sequences in this protocol are always listed in the 5’ -> 3’ orientation. This is the orientation that should be used for ordering.
- We use the standard desalting cleanup option.
- The mammal blocking primer for 18S amplification is formatted for ordering through IDT DNA only.
- If you are using a robot the volume in the plates should be 30 µL per well or greater. 15 µL per well is the absolute minimum.