Dissociation of neuronal culture to single cells for scRNA-seq (10x Genomics) v1

التفاصيل البيبلوغرافية
العنوان: Dissociation of neuronal culture to single cells for scRNA-seq (10x Genomics) v1
المؤلفون: Julie Jerber, James Haldane, Juliette Steer, Daniel Pearce, Minal Patel
بيانات النشر: ZappyLab, Inc., 2020.
سنة النشر: 2020
مصطلحات موضوعية: Chemistry, Biophysics, Genomics, Dissociation (chemistry)
الوصف: This protocol outlines a method for dissociating a human pluripotent stem cell-derived neuronal culture to single cells for loading onto a Chromium 10x chip for single cell RNA-sequencing. This protocol makes a distinction between early neuronal progenitors and mature neuronal cultures, as additional steps and reagents are required in order to sufficiently dissociate the latter. These include: - DNase Vial (D2) - PDS Kit Papain Vial Note: In our labs, iPS cells were undergoing a 52-day long differentiation process to dopaminergic neurons (adapted from doi.org/10.1038/nature10648), and were treated under the 'mature' conditions when harvested from day 20 onward, and as 'early neuronal progenitors' on day 11. Example images of neuronal culture from our labs can be found in the Guidelines of this protocol for reference. This protocol assumes use of: - 12-well tissue culture plates (3.8 cm2 surface area per well ) for samples/wells being harvested for dissociation (see Materials). - A NucleoCounter®NC-200™ for purposes of cell counting.
DOI: 10.17504/protocols.io.bh32j8qe
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::96a66f7515c57abb25c7c597e613a5e9
https://doi.org/10.17504/protocols.io.bh32j8qe
Rights: OPEN
رقم الانضمام: edsair.doi...........96a66f7515c57abb25c7c597e613a5e9
قاعدة البيانات: OpenAIRE
الوصف
DOI:10.17504/protocols.io.bh32j8qe