scholarly journals An extension to: Systematic assessment of commercially available low-input miRNA library preparation kits

RNA Biology ◽  
2020 ◽  
Vol 17 (9) ◽  
pp. 1284-1292
Author(s):  
Fatima Heinicke ◽  
Xiangfu Zhong ◽  
Manuela Zucknick ◽  
Johannes Breidenbach ◽  
Arvind Y.M. Sundaram ◽  
...  
RNA Biology ◽  
2019 ◽  
Vol 17 (1) ◽  
pp. 75-86 ◽  
Author(s):  
Fatima Heinicke ◽  
Xiangfu Zhong ◽  
Manuela Zucknick ◽  
Johannes Breidenbach ◽  
Arvind Y. M. Sundaram ◽  
...  

2020 ◽  
Author(s):  
Fatima Heinicke ◽  
Xiangfu Zhong ◽  
Manuela Zucknick ◽  
Johannes Breidenbach ◽  
Arvind Y.M. Sundaram ◽  
...  

AbstractHigh-throughput sequencing has emerged as the favoured method to study microRNA (miRNA) expression, but biases introduced during library preparation have been reported. To assist researchers choose the most appropriate library preparation kit, we recently compared the performance of six commercially-available kits on synthetic miRNAs and human RNA, where library preparation was performed by the vendors. We hereby supplement this study with data from two further commonly used kits (NEBNext, NEXTflex) whose manufacturers initially declined to participate. As before, performance was assessed with respect to sensitivity, reliability, titration response and differential expression. Despite NEXTflex employing partially-randomised adapter sequences to minimise bias, we reaffirm that biases in miRNA abundance are kit-specific, complicating the comparison of miRNA datasets generated using different kits.


Lab on a Chip ◽  
2017 ◽  
Vol 17 (15) ◽  
pp. 2619-2630 ◽  
Author(s):  
Gordon D. Hoople ◽  
Andrew Richards ◽  
Yan Wu ◽  
Kota Kaneko ◽  
Xiaolin Luo ◽  
...  

Gel-seq enables researchers to simultaneously prepare libraries for both DNA and RNA from 100 cells.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Dimitra Sarantopoulou ◽  
Soon Yew Tang ◽  
Emanuela Ricciotti ◽  
Nicholas F. Lahens ◽  
Damien Lekkas ◽  
...  

Abstract Library preparation is a key step in sequencing. For RNA sequencing there are advantages to both strand specificity and working with minute starting material, yet until recently there was no kit available enabling both. The Illumina TruSeq stranded mRNA Sample Preparation kit (TruSeq) requires abundant starting material while the Takara Bio SMART-Seq v4 Ultra Low Input RNA kit (V4) sacrifices strand specificity. The SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Pico) by Takara Bio claims to overcome these limitations. Comparative evaluation of these kits is important for selecting the appropriate protocol. We compared the three kits in a realistic differential expression analysis. We prepared and sequenced samples from two experimental conditions of biological interest with each of the three kits. We report differences between the kits at the level of differential gene expression; for example, the Pico kit results in 55% fewer differentially expressed genes than TruSeq. Nevertheless, the agreement of the observed enriched pathways suggests that comparable functional results can be obtained. In summary we conclude that the Pico kit sufficiently reproduces the results of the other kits at the level of pathway analysis while providing a combination of options that is not available in the other kits.


protocols.io ◽  
2016 ◽  
Author(s):  
Benjamin Istace ◽  
Anne Friedrich ◽  
L o ◽  
S bastien ◽  
Emilie Payen ◽  
...  

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Abdul Fatah A. Samad ◽  
Nazaruddin Muhammad Ali ◽  
Ismanizan Ismail

In plants, a group of non-coding small RNA (sRNA) has been provento be an important player in regulating gene expression that can govern network of genetic systems. The two major classes of sRNA which are very extensively studied through deep sequencing, microRNA (miRNA) and small-interfering RNA (siRNA) classes, are well documented. However, the isolation method of sRNA differs depending on the type of sample. Here, we demonstrate the miRNA library preparation using two different Small RNA Library preparation kit, Illumina®TruSeq® Small RNA Preparation and NEBNext® Multiplex Small RNA Library Preparation kit on a plant rich in secondary metabolite Persicaria minor using recommended protocol. The result show NEBNext® Multiplex Small RNA Library Preparation kit can recover small RNA better than Illumina®TruSeq® Small RNA Preparation kit. Thus, this study recommended NEBNext® Multiplex Small RNA Library Preparation kit for miRNA library preparation on Persicaria minor.


2015 ◽  
Vol 12 (11) ◽  
pp. v-vi ◽  
Author(s):  
Cassie Schumacher ◽  
Laurie Kurihara ◽  
Kate Cunningham

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