Oligo ordering

9/16/05

1. Download your sequences from NCBI and place in a file to store.

2. Run the sequences through a primer identification program to select sequences with good melting temps and other properties (no primer dimers, etc).

3. Try to select primers that will work in multiple species by checking your sequences against a multispecies alignment (mouse and rat in general).

4. For Q-RT-PCR, try to skip over an intron-exon boundary to avoid detecting contaminating genomic DNA. Follow the instructions in the Q-RT-PCR manuals for good primer design.

5. If you are adding new sequences, type your sequences into the “Westlab oligo database” excel file in the “Westlab database” folder on the “LaCie disk hard drive. Please fill in all information. Print a copy of the revised list and place in the oligo book.

6. Order primers at http://www.idtdna.com .

            name: ••••••

            paswd: ••••••

choose standard DNA oligos, generally 25nmole scale with standard desalting.

7. When your primers arrive, keep a sticker with the sequence in your notebook and place the primer sheets in the oligo book.