[Simone78]

We've not yet tried this, but I'm considering it -- it sure would be nice to barcode during the Tagmentation step rather than the PCR step....if someone tries this, there's a beer in it for you at the next meeting.....

I am not doing ATAC-seq but I might help you anyway. Instead of spending a lot of money for few ul of Tn5 from Epicentre you could just order the plasmid from Addgene (ID: 60240) and make it yourself. Then you could all the adaptors you like!
We have recently published a paper about it, take a look!

http://www.ncbi.nlm.nih.gov/pubmed/25079858

[rbd]

ATACseq Buffer

Does anybody know what the MgCl2 does in the tagmentation buffer? Does it impact chromatin accessibility during ATACseq?

[nucacidhunter]

I would suggest to look at post #8 link in following thread:
http://seqanswers.com/forums/showthread.php?t=42518

[intregen]

It seems to work just fine: If you can see your post-PCR product on a gel, you'll have a fine library (in our experience). Some of the runs have more mtDNA reads than you would expect from, say, DNase-Seq, but this is something many have reported (and is seen in the data from the original paper).

In our experience, the most significant issue is simply making sure that we don't lose our pellet of cells/nuclei during the process.

I should note that we're only using the buffer recipe you linked to wash the cells. The reaction itself still takes place in the commercial buffer. Probably going to stay that way unless someone has a method for making Tn5 independent of the kit.....

Has anyone used the homemade TD buffer for the ATAC reaction and not just for washes? Does it work just as well?

[kchbio]

Has anyone used the homemade TD buffer for the ATAC reaction and not just for washes? Does it work just as well?

Nop, the home-made one does not work as efficient as the TD buffer from Illumina

[Saravanan]

TD buffer recipe

Many thanks, this looks great. I'll update the thread with progress.

Could you share the TD buffer recipe. I couldn't find it.

Many thanks
Saravanan