I use BaseScope 2.5 HD Red quite routinely and I found myself using about 3 liter (1 bottle) of Wash Buffer at every run. Although it is the cheapest component of the kit, it's still quite expensive considering the amount needed.
Has anyone tried to use SSC, TBS-T or another common wash buffer and was there any meaningful difference in the results?
Many thanks
EDIT: Someone suggested me to try SSC 1x buffer (routinely used for home-made RNA-ish protocols). I use to leave my slides overnight in SSC 5x after the probe incubation, has anyone ever tried to use it as a wash buffer as well?
According to this paper: Article RNAscope A Novel in Situ RNA Analysis Platform for Formalin-...
the recipe for the RNAscope wash buffer is (0.1× SSC, 0.03% lithium dodecyl sulfate). That said, I did not have too much luck with RNAscope using just 0.1x SSC as a wash buffer so perhaps the lithium dodecyl sulfate is critical. Also of potential interest, we often use a pH 6 sodium citrate antigen unmasking buffer (Vector labs H3300, diluted 1:100 in ultrapure water) as the wash solution and this seems to give us much better results than when we use the wash buffer that is sold by ACD.
Hi, Yang - sometimes we get the dust/debris as well when we are using the Red kits. In general, we have found that working in a clean area (best case is a biosafety cabinet), keeping the slides covered in a tray with a lid during all incubation steps, and not letting the color reaction go too long, all help to reduce the presence of debris. Also, after you are finished with the color reaction, you can use a detergent wash (we use 0.2% triton in TBS) and this helps a little bit. The main thing you do NOT want to do is use anything that contains any alcohols or organics as they will cause the chromagen to dissolve. Good luck.
- Badges
- Science method