Question: How many cells need to be seeded for RNA-seq analysis
0
gravatar for sacha.bohler
3.4 years ago by
Netherlands
sacha.bohler0 wrote:

Hello everybody,

I am planning a large scale RNA-seq experiment on SH-SY5Y cells, and was wondering how many cells I need to have per sample to extract a sufficient amount of RNA.

Many thanks in advance,

Sacha

rna-seq cell culture • 4.4k views
ADD COMMENTlink modified 2.0 years ago by Biostar ♦♦ 20 • written 3.4 years ago by sacha.bohler0
2
gravatar for jotan
3.4 years ago by
jotan1.2k
Australia
jotan1.2k wrote:

The best thing to do is to ask your sequencing facility.

For a bog-standard PolyA selected RNA-Seq, you want around 1-3 ug of total RNA. However, some facilities have the capacity to go much lower than this (100 ng) if you need to.

The number of cells needed will depend on the cell type but 1-2 e5 is probably plenty if you're aiming for the lower limit (1 ug). If your cells are easy to grow then seed more and extract more RNA and then just give the sequencing facility as much as they need. 

ADD COMMENTlink written 3.4 years ago by jotan1.2k
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