Enzymatic methyl sequencing detects DNA methylation at single-base resolution from picograms of DNA

亚硫酸氢盐 亚硫酸氢盐测序 DNA甲基化 5-甲基胞嘧啶 甲基化DNA免疫沉淀 生物 DNA 结扎测序 基因组文库 照明菌甲基化试验 基因组DNA 计算生物学 DNA测序 Illumina染料测序 DNA纳米球测序 遗传学 分子生物学 基因 基序列 基因表达
作者
Romualdas Vaisvila,V. K. Chaithanya Ponnaluri,Zhiyi Sun,Bradley W. Langhorst,Lana Saleh,Shengxi Guan,Nan Dai,Matthew A. Campbell,Brittany S. Sexton,Katherine Marks,Mala Samaranayake,James C. Samuelson,Heidi E. Church,Esta Tamanaha,Ivan R. Corrêa,Sriharsa Pradhan,Eileen T. Dimalanta,Thomas C. Evans,Louise Williams,Theodore B. Davis
出处
期刊:Genome Research [Cold Spring Harbor Laboratory]
卷期号:31 (7): 1280-1289 被引量:163
标识
DOI:10.1101/gr.266551.120
摘要

Bisulfite sequencing detects 5mC and 5hmC at single-base resolution. However, bisulfite treatment damages DNA, which results in fragmentation, DNA loss, and biased sequencing data. To overcome these problems, enzymatic methyl-seq (EM-seq) was developed. This method detects 5mC and 5hmC using two sets of enzymatic reactions. In the first reaction, TET2 and T4-BGT convert 5mC and 5hmC into products that cannot be deaminated by APOBEC3A. In the second reaction, APOBEC3A deaminates unmodified cytosines by converting them to uracils. Therefore, these three enzymes enable the identification of 5mC and 5hmC. EM-seq libraries were compared with bisulfite-converted DNA, and each library type was ligated to Illumina adaptors before conversion. Libraries were made using NA12878 genomic DNA, cell-free DNA, and FFPE DNA over a range of DNA inputs. The 5mC and 5hmC detected in EM-seq libraries were similar to those of bisulfite libraries. However, libraries made using EM-seq outperformed bisulfite-converted libraries in all specific measures examined (coverage, duplication, sensitivity, etc.). EM-seq libraries displayed even GC distribution, better correlations across DNA inputs, increased numbers of CpGs within genomic features, and accuracy of cytosine methylation calls. EM-seq was effective using as little as 100 pg of DNA, and these libraries maintained the described advantages over bisulfite sequencing. EM-seq library construction, using challenging samples and lower DNA inputs, opens new avenues for research and clinical applications.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
2秒前
2秒前
3秒前
赘婿应助fifteen采纳,获得10
4秒前
搜集达人应助稳重驳采纳,获得10
4秒前
yinxx发布了新的文献求助10
5秒前
zzp完成签到,获得积分10
5秒前
灯火完成签到,获得积分10
6秒前
肥蛇外传完成签到 ,获得积分10
6秒前
麦尔哈巴完成签到 ,获得积分10
7秒前
wg发布了新的文献求助30
8秒前
xiaoxiang_1001完成签到,获得积分10
10秒前
赘婿应助wny采纳,获得10
11秒前
11秒前
11秒前
王唯任完成签到,获得积分10
12秒前
14秒前
1點點cui完成签到,获得积分10
15秒前
Hello应助不爱胡椒采纳,获得10
15秒前
会游泳的思维应助罗是一采纳,获得10
16秒前
背后归尘完成签到,获得积分10
16秒前
1點點cui发布了新的文献求助10
18秒前
19秒前
93发布了新的文献求助10
20秒前
罗是一完成签到,获得积分10
22秒前
rong应助如沐采纳,获得10
23秒前
24秒前
Yangzx发布了新的文献求助10
25秒前
suuummmer完成签到,获得积分10
27秒前
怎么说应助1點點cui采纳,获得10
27秒前
30秒前
31秒前
桐桐应助saikun采纳,获得10
31秒前
Executor完成签到,获得积分10
31秒前
风趣丝发布了新的文献求助10
34秒前
Jiang应助zz采纳,获得20
34秒前
34秒前
35秒前
小冉发布了新的文献求助10
37秒前
高分求助中
좌파는 어떻게 좌파가 됐나:한국 급진노동운동의 형성과 궤적 2500
Sustainability in Tides Chemistry 1500
TM 5-855-1(Fundamentals of protective design for conventional weapons) 1000
CLSI EP47 Evaluation of Reagent Carryover Effects on Test Results, 1st Edition 800
Cognitive linguistics critical concepts in linguistics 800
Threaded Harmony: A Sustainable Approach to Fashion 799
Livre et militantisme : La Cité éditeur 1958-1967 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3053642
求助须知:如何正确求助?哪些是违规求助? 2710842
关于积分的说明 7423746
捐赠科研通 2355391
什么是DOI,文献DOI怎么找? 1247143
科研通“疑难数据库(出版商)”最低求助积分说明 606239
版权声明 595992