Agarose amplification based sequencing characterization cell-free RNA in preimplantation spent embryo medium

化学 胚泡 核糖核酸 胚胎 琼脂糖 分子生物学 RNA提取 细胞生物学 色谱法 生物化学 基因 胚胎发生 生物
作者
Huajuan Shi,Qinyu Ge,Min Pan,Yuqi Sheng,Ting Qi,Ying Zhou,Yuqing Sun,Yunfei Bai,Lingbo Cai
出处
期刊:Analytica Chimica Acta [Elsevier BV]
卷期号:1296: 342331-342331
标识
DOI:10.1016/j.aca.2024.342331
摘要

The cell-free RNA (cf-RNA) of spent embryo medium (SEM) has aroused a concern of academic and clinical researchers for its potential use in non-invasive embryo screening. However, comprehensive characterization of cf-RNA from SEM still presents significant technical challenges, primarily due to the limited volume of SEM. Hence, there is urgently need to a small input liquid volume and ultralow amount of cf-RNA library preparation method to unbiased cf-RNA sequencing from SEM. (75) Here, we report a high sensitivity agarose amplification-based cf-RNA sequencing method (SEM-Acf) for human preimplantation SEM cf-RNA analysis. It is a cf-RNA sequencing library preparation method by adding agarose amplification. The agarose amplification sensitivity (0.005 pg) and efficiency (105.35 %) were increased than that of without agarose addition (0.45 pg and 96.06 %) by ∼ 90 fold and 9.29 %, respectively. Compared with SMART sequencing (SMART-seq), the correlation of gene expression was stronger in different SEM samples by using SEM-Acf. The cf-RNA number of detected and coverage uniformity of 3′ end were significantly increased. The proportion of 5′ end adenine, alternative splicing events and short fragments (<400 bp) were increased. It is also found that 4‐mer end motifs of cf-RNA fragments was significantly differences between different embryonic stage by day3 spent cleavage medium and day5/6 spent blastocyst medium. (141) This study established an efficient SEM amplification and library preparation method. Additionally, we successfully described the characterizations of SEM cf-RNA in preimplantation embryo using SEM-Acf, including expression features and fragment lengths. SEM-Acf facilitates the exploration of cf-RNA as a noninvasive embryo screening biomarker, and opens up potential clinical utilities of small input liquid volume and ultralow amount cf-RNA sequencing. (59).
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
PDF的下载单位、IP信息已删除 (2025-6-4)

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
scxl2000完成签到 ,获得积分10
刚刚
1秒前
轻轻完成签到,获得积分20
1秒前
董湘澜完成签到,获得积分10
1秒前
量子星尘发布了新的文献求助10
2秒前
Ronnie发布了新的文献求助10
3秒前
YJJ完成签到,获得积分10
4秒前
打打应助likes采纳,获得10
5秒前
左囧发布了新的文献求助10
5秒前
艺玲发布了新的文献求助10
6秒前
6秒前
俞圣爱发布了新的文献求助10
8秒前
斯文败类应助穆晴筱筱采纳,获得10
13秒前
富含亮氨酸重复应助艺玲采纳,获得10
13秒前
烂漫平蓝发布了新的文献求助50
13秒前
16秒前
16秒前
17秒前
17秒前
赖晨靓完成签到 ,获得积分10
18秒前
zhangmeimei完成签到,获得积分10
18秒前
JTYJRTY发布了新的文献求助10
19秒前
在水一方应助学术悍匪采纳,获得10
20秒前
不钓鱼发布了新的文献求助10
20秒前
乐乐应助会飞的野马采纳,获得10
21秒前
梦溪发布了新的文献求助10
22秒前
overThat完成签到,获得积分10
22秒前
YamDaamCaa应助yiqi采纳,获得30
23秒前
彩色不评完成签到,获得积分10
24秒前
刀笔吏完成签到,获得积分10
26秒前
Hiker完成签到,获得积分10
27秒前
28秒前
科研通AI5应助xuchen采纳,获得10
29秒前
勤奋高丽发布了新的文献求助20
30秒前
30秒前
ecoli完成签到,获得积分20
32秒前
33秒前
likes发布了新的文献求助10
34秒前
学术悍匪发布了新的文献求助10
34秒前
村医完成签到,获得积分10
35秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 700
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
热门求助领域 (近24小时)
化学 材料科学 医学 生物 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 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3976253
求助须知:如何正确求助?哪些是违规求助? 3520405
关于积分的说明 11203301
捐赠科研通 3257028
什么是DOI,文献DOI怎么找? 1798589
邀请新用户注册赠送积分活动 877755
科研通“疑难数据库(出版商)”最低求助积分说明 806521