TARGET-Seq: A Protocol For High-Sensitivity Single-Cell Mutational Analysis and Parallel RNA Sequencing

Published: 10-09-2020| Version 1 | DOI: 10.17632/k92cnf2fph.1
Contributor:
Alba Rodriguez-Meira

Description

TARGET-Seq is a protocol for TARGETed high-sensitivity single-cell mutational analysis, parallel RNA SEQuencing and cell-surface proteomics, with extremely low allelic-dropout rates . In this manuscript, we present a detailed step-by-step protocol for the application of TARGET-seq (Rodriguez-Meira et al, Molecular Cell, 2019), including troubleshooting tips, approaches for automation and methods for high-throughput multiplexing of libraries. In this dataset, we provide program files for specific liquid handlers corresponding to each step of the protocol.

Files

Steps to reproduce

Supplemental Methods Methods S1, related to Step 1.2 Point 13. Lysis buffer dispending program for the MANTIS liquid handler platform. Methods S2, related to Step 2.1 Point 36. RT buffer dispending program for the MANTIS liquid handler platform. Methods S3, related to Step 2.2 Point 40. PCR buffer dispending program for the MANTIS liquid handler platform. Methods S4, related to Step 2.3 Point 48. Biomek FxP program used for cDNA pooling and genotyping stock preparation. Methods S5, related to Step 3 Point 54. Biomek FxP program used for targeted library preparation PCR1 genotyping. Methods S6, related to Step 3 Point 58. Biomek FxP program used for barcode aliquoting. Methods S7, related to Step 3 Point 61. Biomek FxP program used for targeted library preparation PCR2 genotyping. Methods S8, related to Step 3 Point 62. Biomek FxP program used for genotyping pooling.