JoVE Journal

Genetics

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Nanyang Technological University

Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples

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11.9K Views

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07:30 min

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June 8th, 2020

DOI :

10.3791/61060-v

June 8th, 2020

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Yelena Levin*¹, Keyur Talsania*¹^,², Bao Tran¹, Jyoti Shetty¹, Yongmei Zhao¹^,², Monika Mehta¹

¹NCI CCR Sequencing Facility, Frederick National Laboratory for Cancer Research, ²Advanced Biomedical and Computational Sciences, Frederick National Laboratory for Cancer Research

Podsumowanie

This method describes the steps to improve the quality and quantity of sequence data that can be obtained from formalin-fixed paraffin-embedded (FFPE) RNA samples. We describe the methodology to more accurately assess the quality of FFPE-RNA samples, prepare sequencing libraries, and analyze the data from FFPE-RNA samples.

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FFPE RNA Samples

Gene Expression Data

RNA Quality Assessment

Characterization

Sequencing Data

Library Preparation

Bioanalysis Software

RNA Fragment Distribution

Illumina Experiment Manager

Sample Library Preparation

PhiX Control Library

Sequencing Reagent Kits

Flow Cell Preparation

Rozdziały w tym wideo

0:04

Introduction

1:21

RNA Quantity and Quality Assessment

2:03

Sequencing

4:01

Data Analysis and Quality Assessment

4:33

Results: Representative Degraded Formalin-Fixed Paraffin Embedded (FFPE)- RNA Sample Sequencing and Analysis

6:28

Conclusion

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