Ok, we did try @ParseBio SPLIT-Seq WT kit. We don’t have the data yet but workflow-wise it was super easy. I really liked the simplicity and that the kit comes with pretty much all you need. Totally manageable for anybody who has done 96-well work and multi-channel pipettes. 1/6
They also have a super easy to fill out Excel sheet that makes all the calculations for you. Peace of mind right there. If you counted your cells correctly then you are for a good start. 2/6
You DO need at least 3x more cells input to ensure you end up with the targeted numbers of cells at the end. Of course, viability and cell numbers before and after fixation is critical. 3/6
Split and pooling is just like any other approach of the likes.
Sublibraries (up to 8) prep is standard. The Userguide is VERY comprehensive which is great. cDNA/Lib QC will be more expensive as you now have 8 cDNA/Libs to check instead of 1. But, sublibraries are useful! 4/6
Sequencing-wise, well…it will be more costly than other techs because you need a 200 cycles kit. R1: 74 + R2: 86 = 160 cycles needed. #NovaSeq 100 cycles kit is worth only 138, so a 200 cycles kit is needed. 5/6 @illumina
Probably the most important part, Parse’s support team was super helpful…in particular Anna Malinkevich.
All in all, I think it’s a great product. Great start @ParseBio! 6/6
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Assay for Transposase-Accessible Chromatin (ATAC-Seq) was originally published in in 2013 (@naturemethods PMID: 24097267) by @JD_Buenrostro in the labs of H. Chang and W. Greenleaf at @Stanford.
ATAC-Seq is a VERY efficient technique, and was quickly implemented at #singlecell level in 2015 by the same team (@nature PMID: 26083756). These are good reads and help understand why this method is so revolutionary.