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Method Article
Genomic DNA Isolation from Tissue Samples and Library Prep for Tuba-Seq Barcode Analysis
Zoe Rogers
Winslow Lab
Corresponding Author
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This protocol should be used in preparation from tumor barcode sequencing (Tuba-seq) with a goal of directly quantifying the tumor sizes within tissues. This protocol could also be used to quantify cells from culture, but here we only discuss genomic DNA extraction from whole tissues. The protocol begins with phenol-chloroform gDNA extraction of homogenized tissue. Once gDNA has been harvested, the barcode containing region is amplified with custom primers which allow attachment of Illumina adapters in a single step. Finally, we discuss how to pool samples before sending your library in for sequencing. The entire process takes 3-4 days. This protocol accompanies Rogers et al, Nature Methods, published online May 22, 2017 (nmeth.4297).
Keywords
Tuba-seq, barcoding, tumor suppressors, cancer
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This is a preprint. It has not completed peer review.
Method Article
Genomic DNA Isolation from Tissue Samples and Library Prep for Tuba-Seq Barcode Analysis
Zoe Rogers
Winslow Lab
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 25 May, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol should be used in preparation from tumor barcode sequencing (Tuba-seq) with a goal of directly quantifying the tumor sizes within tissues. This protocol could also be used to quantify cells from culture, but here we only discuss genomic DNA extraction from whole tissues. The protocol begins with phenol-chloroform gDNA extraction of homogenized tissue. Once gDNA has been harvested, the barcode containing region is amplified with custom primers which allow attachment of Illumina adapters in a single step. Finally, we discuss how to pool samples before sending your library in for sequencing. The entire process takes 3-4 days. This protocol accompanies Rogers et al, Nature Methods, published online May 22, 2017 (nmeth.4297).