Is possible to stain with SYBR green

[mx1970]

We are going to buy a transilluminator to cut the slide from the gel to make the library and We are thinking in get a transiluminator of blue light instead UV.

Could be possible to stain the DNA with SYBR green, purify the DNA and make the library with this DNA without problems in the sequencing?
Thanks

[ajthomas]

It's a good idea to use a blue transilluminator rather than UV because the UV light will damage the DNA very rapidly.

I don't know about SYBR green, but Invitrogen sells a dye called Sybr Safe with is for exactly what you are talking about doing. My lab switched to it several years ago for all of our gels and haven't looked back since. Invitrogen even sells a blue transilluminator for it.

[twaddlac]

I believe it is safe, but my lab hasn't done it. I did come across this which does say that they specifically tested the effects of SYBR green on sequencing and it did not suggest interference, though they used a sequencing kit.

http://www.epibio.com/newsletter/11_4_17-19.pdf

If you're going for NGS then I would be highly skeptical since the purest DNA yields plenty of sequencing on its own.

Hope this helps!

[basilash]

We are going to buy a transilluminator to cut the slide from the gel to make the library and We are thinking in get a transiluminator of blue light instead UV.

Could be possible to stain the DNA with SYBR green, purify the DNA and make the library with this DNA without problems in the sequencing?
Thanks

Yes it is quite OK: I am using a blue transilluminator and SYBR green stained gels for NGS libraries. It is much more safe and convenient as compared to using UV.

[bbeitzel]

We use a blue light transilluminator ( the Dark Reader, http://www.clarechemical.com/ , thought I'd give them a shout out since they were the original) with GelStar stain (http://tinyurl.com/7bdzkl8) for all of our manual size selections for NGS. GelStar is a very sensitive stain on the Dark Reader (probably 5-10X more sensitive than ethidium bromide) , so you have to be careful not to overload the gels.