Resources

If you are interested in the Single Cell Core, please start by completing an application and wait to be contacted by the staff to discuss the scope of your request. 

NOTE the SCC does not provide sequencing or anaysis as a service. 

Data the SCC collected on both inDrops and 10x instruments is available here:

https://www.synapse.org/#!Synapse:syn17017240/wiki/584708

For full service bioinformatic analysis service we suggest you work with the Chan Bioinformatics Core.

http://bioinformatics.sph.harvard.edu/

Sample Requirements/Tips

Number of cells: 25,000 – 30,000 at a minimum. 
Cells should be at a density of more than 100,000 cells/ml in at least 150ul.

Viability: Ideally greater than 90%

Buffer conditions: Buffer should NOT contain Mg2+, Ca2+ or EDTA.
Most of users resuspend their cells in PBS (without Mg2+, Ca2+ or EDTA) + BSA (upto 1%)

  • Single-nucleus RNA sequencing can be run with inDrops. Several users perform this routinely.
  • A standard inDrops day encapsulates 6,000 cells per sample for up to 6 samples.
  • Optiprep information
  • CITE-seq / Cell Hashing have succesfully run on both inDrops and 10x.

10x Single-cell sample prep resources

10x Genomics provides a tremendous amount of sample prep protocol advice.  Most of the sample prep requirements for 10x are the same for inDrops.  This is a great resource to use for sample prep optimization.

Protocols.io

There are a variety of scRNA-Seq and scNuc-Seq methods available.

Click here for a search of "single cell dissociation scRNA-Seq"

InDrops Method Publications

Sequencing Guidelines

Most users sequence their libraries on a NextSeq 75bp run:

Read 1: 61 
Read 2: 14
Index 1: 8
Index 2: 8

It is common to have 70-80% valid reads after filtering for valid cell barcodes.

Sequencing on a their libraries on a NovaSeq 100bp run:

Read 1: 86 
Read 2: 14
Index 1: 8
Index 2: 8

Our initial experience with inDrops runs on a NovaSeq indicate that we are having some trouble on the Index reads. However, we think we have identified the issue being that the NovaSeq runs of inDrops require that you recieve UN-TRIMMED Fastq files. Adapter trimming seems to be causing some loss of valid reads causeing you go only get 6 bp of some of the 8bp index read. However the un-trimmed fastq files does fix this issue.  Please let us know if you experience any issues on a NovaSeq run so we can help troubleshoot and understand the issues for all inDrops users.

library structure

Policies

  • Full day or half day services are available. To qualify as a half day run, you must have 3 or fewer samples and be collecting no more than 15,000 cells in total.
  • Run preparation begins 1 hour in advance of the scheduled time. To avoid reagent and service fees, please notify us of any delays or cancellations at least 2 hours in advance of your start time.
  • If you are more than an hour late for your scheduled run, the SCC staff reserves the right to cancel your run and charge you for the reserved time and reagents used in preparation for your run.
  • The SCC will hold on to backup encapsulated cells for 1 year after your run date. If you need your backup emulsion please retrieve it from the  SCC within a few months of your run.