Investigating the Influences of Random-Region Length on Aptamer Selection Efficiency Based on Capillary Electrophoresis–SELEX and High-Throughput Sequencing

Author(s):  
Chao Zhu ◽  
Linsen Li ◽  
Ge Yang ◽  
Feng Qu
2018 ◽  
Author(s):  
Karlis Pleiko ◽  
Liga Saulite ◽  
Vadims Parfejevs ◽  
Karlis Miculis ◽  
Egils Vjaters ◽  
...  

ABSTRACTAptamers have in recent years emerged as a viable alternative to antibodies. High-throughput sequencing (HTS) has revolutionized aptamer research by increasing the number of reads from a few (using Sanger sequencing) to millions (using an HTS approach). Despite the availability and advantages of HTS compared to Sanger sequencing, there are only 50 aptamer HTS sequencing samples available on public databases. HTS data in aptamer research are primarily used to compare sequence enrichment between subsequent selection cycles. This approach does not take full advantage of HTS because the enrichment of sequences during selection can be due to inefficient negative selection when using live cells. Here, we present a differential binding cell-SELEX (systematic evolution of ligands by exponential enrichment) workflow that adapts theFASTAptamertoolbox and bioinformatics tooledgeR, which are primarily used for functional genomics, to achieve more informative metrics about the selection process. We propose a fast and practical high-throughput aptamer identification method to be used with the cell-SELEX technique to increase the aptamer selection rate against live cells. The feasibility of our approach is demonstrated by performing aptamer selection against a clear cell renal cell carcinoma (ccRCC) RCC-MF cell line using the RC-124 cell line from healthy kidney tissue for negative selection.


BioTechniques ◽  
2011 ◽  
Vol 51 (6) ◽  
Author(s):  
Shawn Hoon ◽  
Bin Zhou ◽  
Kim Janda ◽  
Sydney Brenner ◽  
Jonathan Scolnick

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Julissa J. Sánchez-Velásquez ◽  
Lorenzo E. Reyes-Flores ◽  
Carmen Yzásiga-Barrera ◽  
Eliana Zelada-Mázmela

Abstract Objective The advancement of molecular techniques in an era in which high-throughput sequencing has revolutionized biology renders old-fashioned alternatives to high-throughput methods obsolete. Such advanced molecular techniques, however, are not yet accessible to economically disadvantaged region-based laboratories that still obtain DNA profiles using gel-based techniques. To explore whether cost-efficient techniques can produce results that are as robust as those obtained using high-throughput methods, we compared the performance of polyacrylamide gel electrophoresis (PAGE)- and capillary electrophoresis (CE)-derived genomic data in estimating genetic diversity and inferring relatedness using 70 individuals of fine flounder (Paralichthys adspersus) selected from a hatchery population and genotyped for five microsatellite loci. Results Here, we show that PAGE- and CE-derived genomic datasets yield comparable genetic diversity levels regarding allelic diversity measures and heterozygosity. However, relatedness inferred from each dataset showed that the categorization of dyads in the different relationship categories strongly differed. This suggests that while scientists can reliably use PAGE-derived genomic data to estimate genetic diversity, they cannot use the same for parentage testing. The findings could help laboratories committed to population research not be discouraged from using the PAGE system if high-throughput technologies are unavailable and the method is adequate to address the biological question.


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