Griffitts:PCR primers

General

 * Primers should generally be 20–22 nt long with 10–12 Gs and Cs
 * Remember to use the reverse compliment for your reverse primer

Sequencing

 * Design one forward primer for every ~500 bp
 * You won't get good sequence where your primer sits, so design your first primer to be a little upstream from your sequence
 * You only need one reverse primer

Cloning

 * Check for restriction sites within your target before adding them to your primers
 * Add a "CGC" to the ends of your primers

Gene Deletion

 * Check for restriction sites within your target before adding them to your primers
 * Add a "CGC" to the ends of your primers
 * Clone ~500 bp upstream and downstream of the gene you're intending to delete and include ~20 bp of the target gene
 * For your internal primers, give them 10-12 bp homology with each other for Overlap-extension PCR

Primer Hydration

 * Find the nmoles on the tube (e.g. 24.65 nmoles)
 * Multiply by ten (e.g. 24.65 × 10 = 246.5)
 * Take that number and add that many μL of TE buffer (e.g. add 240 to 250 μL)
 * This will give you 100X primers