In article <326E6224.5BBC at vetsci.microvet.arizona.edu>,
billing at vetsci.microvet.arizona.edu wrote:
> Is anyone aware of a stain that is specific for single stranded DNA? I
> seem to recall something about acridine orange, but the ol' brain cells
> can't be certain.
>> I am trying to find an easy method to detect single stranded
> plasmid intermediates on agarose gels? If not specific stains, any
> other ideas?
>> TIA
>> Steve
Hi, Steve,
when viewed on a UV transilluminator (around 300 nm), acridine orange will
stain single stranded DNA bright orange-reddish, and double stranded DNA
yellowish green. The protocol is: (from USB T7-GEN in vitro mutagenesis
kit)
- run your gel (TAE or TBE)
- soak in a solution of 30 microgram/ml acridine orange in 10 mM sodium
phosphate pH 7.0, for 15-30 min at room temp (protect from light)
- destain very thoroughly, to reduce background, by several changes of HOT
tap water over 15-20 min. Works well for me...
Good luck,
Regina
Recombinant Protein Expression Core/ICBR
