# Single Cell Sequencing ## Cell Isolation Techniques Cells must first be compartmentalized or isolated In order for cells within a sample to be analyzed on a single cell basis, they must first be - [[Microfluidics]] - Spatial - Compartmentalization - Signal Association: some techniques avoid having to physically isolate cells by repeatedly sending out electronic signals and reading the response from the sample ## Examples of Single Cell Sequencing ### scATAC-Seq ### scRNA-Seq Sequencing DNA can be useful for studying properties such as the conservation of genes across species, but when it comes to **what genes are being expressed** its important to focus on the **RNA present within cells**. [[Transcription|Transcription factors]] > In an ideal world we would simply measure the presence of the proteins themselves but the technology required to do that is still very limited ![[Pasted image 20230107015414.png|300]] ## Drawbacks of Single Cell Sequencing ### Classifying Cell Types ## Data Normalization [RPKM, FPKM and TPM, Clearly Explained!!!](https://www.youtube.com/watch?v=TTUrtCY2k-w) - Analyzing longer, single strands of RNA vs spliced separate strands - Total sequencing depth (how much RNA is in each sample) - Gene length variation (how long is each gene) ### Single Cell RNA-Seq ### Bulk RNA-Seq