The Effect of Elution Volume for Immunoprecipitation on m6A-Seq Analysis

² R&D Department, LC-Bio Techno2R&D Department, LC-Bio Technology Co., Ltd, Hangzhou, Zhejiang 310018, Chinalogy Co., Ltd, Hangzhou, Zhejiang 310018, China,

³ College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China,

PAR 2021 , 5(4), 1–5; https://doi.org/10.26689/par.v5i4.2239

© 2021 by the Authors. Licensee Whioce Publishing, Singapore. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

Abstract

Objective: To develop a cost-effective method to reduce the time consumption of elution in immunoprecipitation. Method: Two volumes (125 ?L for Group C and 100 ?L for Group T) of elution buffer were used to explore whether smaller volume could save testing time. Result: Time consumption of elution in Group T was significantly shorter than that in Group C, while the efficiency of eluted m⁶A-containing fragments and the performance of m⁶A-Seq as indicated by m⁶A peak distributions showed no difference between the two groups. Conclusion: A smaller volume of elution buffer was an economical way to reduce time consumption in immunoprecipitation.

References

Jiang XL, Liu BY, Nie Z, et al., 2021, The Role of m6A Modification in the Biological Functions and Diseases. Signal Transduction and Targeted Therapy, 6(1): 74.

Meyer KD, Saletore Y, Zumbo P, et al., 2012, Comprehensive Analysis of MRNA Methylation Reveals Enrichment in 3’ UTRS and Near Stop Codons. Cell, 149(7): 1635-1646.

Dominissini D, Moshkovitz S, Schwartz, et al., 2012, Topology of the Human and Mouse m6a RNA Methylomes Revealed by m6a-Seq. Nature, 485(7397): 201-206.

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