Genomic DNA Amplification and Sequencing

Genomic DNA was extracted from sorted hematopoietic cells and amplified using Phusion PCR Master Mix (Thermo Fisher Scientific, #F-531 L). PCR reactions were halted once they had progressed halfway through the exponential phase. The PCR product was purified and analyzed using an Illumina NextSeq 500 at the USC Epigenome Center. Sequencing data were analyzed, as previously described (37 (link)–39 (link)). The lentiviral vector that delivers genetic barcodes also conveys GFP expression that marks barcoded cells.

Free full text: Click here

Jiang D., Chowdhury A.Y., Nogalska A., Contreras J., Lee Y., Vergel-Rodriguez M., Valenzuela M, & Lu R. (2024). Quantitative association between gene expression and blood cell production of individual hematopoietic stem cells in mice. Science Advances, 10(4), eadk2132.

Publication 2024

Phusion PCR Master Mix NextSeq 500

Corresponding Organization : Broad Center

Top 5 similar protocols

Variable analysis

independent variables

Extraction and amplification of genomic DNA from sorted hematopoietic cells

dependent variables

Sequencing data from the Illumina NextSeq 500 analysis

control variables

Phusion PCR Master Mix (Thermo Fisher Scientific, #F-531 L) used for PCR amplification
PCR reactions halted once they had progressed halfway through the exponential phase
Sequencing data analyzed as previously described (references 37, 38, and 39)

controls

The lentiviral vector that delivers genetic barcodes also conveys GFP expression, which marks barcoded cells

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!