SYBR Green staining is not something that we routinely do. It stains dsDNA, and can be used to check whether polony amplification worked without going through a single or multi-base sequencing protocol. However, the signal-to-noise ratio is so poor, and the images so messy, that we generally prefer performing either a hybridization with a Cy5 or Cy3 labeleld primer or performing a single-base-extension even for simply checking how well a polony amplification worked. Nevertheless, this protocol can be useful for trouble-shooting and certain applications.
1. Mix 8 uL of concentrated SYBR Green (10000x) into 50 mL 0.5X TBE.
2. Incubate slides with mix in plastic Coplin jar on shaker for 20 minutes.
3. Wash once with 0.5X TBE (20 minutes on shaker in plastic Coplin jar).