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Series GSE174280 Query DataSets for GSE174280
Status Public on Mar 23, 2022
Title Extensive evaluation of ATAC-seq protocols for native or formaldehyde-fixed nuclei
Organisms Homo sapiens; Mus musculus
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The "Assay for Transposase Accessible Chromatin sequencing" (ATAC-seq) is an efficient and easy to implement protocol to measure chromatin accessibility that has been widely used in multiple applications studying gene regulation. While several modifications or variants of the protocol have been published since it was first described, there has not yet been an extensive evaluation of the effects of specific protocol choices head-to-head in a consistent experimental setting. In this study, we tested multiple protocol options for major ATAC-seq components (including three reaction buffers, two reaction temperatures, two enzyme sources, and the use of either native or fixed nuclei) in a well-characterized cell line. In addition, the native conditions were tested in a primary sample type (mouse lung tissue) with two different input amounts. In general, native samples yielded more peaks (particularly at loci not overlapping transcription start sites) than fixed samples, and the temperature at which the enzymatic reaction was carried out had a major impact on data quality metrics for both fixed and native nuclei. However, the effect of various conditions tested was not always consistent between the native and fixed samples. For example, the Nextera and Omni buffers were largely interchangeable across all other conditions, while the THS buffer resulted in markedly different profiles in native samples. In-house and commercial enzymes performed similarly. We found that the relationship between commonly used measures of library quality differed across temperature and fixation, and so evaluating multiple metrics in assessing the quality of a sample is recommended. Notably, we also found that these choices can bias the functional class of elements profiled and so we recommend evaluating several formulations in any new experiments. Finally, we hope the ATAC-seq workflow formulated in this study on crosslinked samples will help to profile archival clinical specimens.
 
Overall design We created 24 experimental conditions with all possible combinations of three ATAC-seq buffers, two reaction temperatures, two Tn5 enzyme sources, and the use of either native or fixed cells in GM12878 cell line. A total of 96 samples were generated with four replicates for each condition. All experimental conditions were also repeated in native nuclei isolated from lungs from three different mice, and performed using two different input amounts. Through this extensive comparison, we were able to observe the effect of different ATAC-seq conditions on data quality and to examine the utility and potential redundancy of various quality metrics.
 
Contributor(s) Zhang H, Rice ME, Alvin JW, Farrera-Gaffney D, Galligan JJ, Johnson ML, Cusanovich DA
Citation(s) 35296236
Submission date May 11, 2021
Last update date Mar 31, 2022
Contact name Hao Zhang
E-mail(s) haozhang1@arizona.edu
Organization name The University of Arizona
Department CMM
Lab Cusanovich
Street address 1230 N Cherry Ave
City Tucson
State/province AZ
ZIP/Postal code 85719
Country USA
 
Platforms (2)
GPL21626 NextSeq 550 (Mus musculus)
GPL21697 NextSeq 550 (Homo sapiens)
Samples (168)
GSM5290833 GM12878-Native-20k-Nextera_TD-37C-Nextera_Tn5-Rep1
GSM5290834 GM12878-Native-20k-Omni_TD-37C-Nextera_Tn5-Rep1
GSM5290835 GM12878-Native-20k-THS_TD-37C-Nextera_Tn5-Rep1
Relations
BioProject PRJNA729079
SRA SRP319568

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MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE174280_RAW.tar 70.4 Mb (http)(custom) TAR (of NARROWPEAK)
GSE174280_dev_matrix.txt.gz 337.5 Kb (ftp)(http) TXT
GSE174280_readcounts.txt.gz 10.5 Mb (ftp)(http) TXT
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