RNA was extracted from lin-CD34+, lin-CD34+CD38-CD90+CD93- and lin-CD34+CD38-CD90+CD93+ populations, reverse transcribed, and 14 cycles of gene-specific amplification performed using the Applied Biosystem pre-amplification kit with relevant primer sets. Following amplification, 2uL of the resultant product was used for multiplex PCR reaction, as described 25 (link). The rest of the resultant products were loaded in triplicate onto pre-primed 96x96 Fluidigm microfluidic dynamic arrays and analyzed according to the manufacturer’s instructions. To assess single cell gene expression, Fluidigm C1™ was used. Analysis was performed using R 3.3.3 under macOS 10.13.2. Data processing and normalization were performed independently for each chip. Only those genes with detectable expression in at least 10 CD93+ and 10 CD93- cells within the same chip were analyzed. All the expression values were normalized using the –ΔΔCt method 24 (link).
Fluidigm microfluidic dynamic arrays
Manufactured by Standard BioTools
The Fluidigm microfluidic dynamic arrays are a type of lab equipment designed for high-throughput, multiplex analysis of biological samples. The core function of these arrays is to enable the simultaneous measurement of multiple analytes or targets within a single sample, facilitating efficient and comprehensive data collection.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fluidigm microfluidic dynamic arrays
1
Single-cell gene expression analysis of hematopoietic progenitors
2
Transcriptomic Analysis of Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was extracted from lin−CD34+, lin−CD34+CD38−CD90+CD93−, and lin−CD34+CD38−CD90+CD93+ populations, reverse transcribed, and 14 cycles of gene-specific amplification performed using the Applied Biosystem pre-amplification kit with relevant primer sets. Following amplification, 2 uL of the resultant product was used for multiplex PCR reaction, as described [25 (link)]. The rest of the resultant products were loaded in triplicate onto pre-primed 96 × 96 Fluidigm microfluidic dynamic arrays and analyzed according to the manufacturer’s instructions. To assess single-cell gene expression, Fluidigm C1™ was used. Analysis was performed using R 3.3.3 under macOS 10.13.2. Data processing and normalization were performed independently for each chip. Only those genes with detectable expression in at least 10 CD93+ and 10 CD93− cells within the same chip were analyzed. All the expression values were normalized using the –ΔΔCt method [24 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!