Amplicon Library
Amplicon Library - Related Videos
08:57
Amplicon Sequencing using the Long-Read Sequencing Technologies
0 Views •
2025
This protocol was optimized for targeted deep sequencing of 18 drug-resistance regions in Mycobacterium tuberculosis using a long-read sequencing platform, followed by analysis with a tuberculosis-specific bioinformatics pipeline designed for long-read data.
10:24
Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
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Cited by 119 •
2014
Characterizing microbial community has been a longstanding goal in environmental microbiology. Next-generation sequencing methods now allow for the characterization of microbial communities at an unprecedented depth with minimal cost and labor. We detail here our approach to sequence bacterial 16S ribosomal RNA genes using a benchtop sequencer.
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 11 •
2011
Here we use a human LentiPlex pooled library and traditional sequencing methods to identify gene targets promoting cell survival. We demonstrate how to set up and deconvolute a LentiPlex screen and validate the results.
10:32
A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
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Cited by 2 •
2017
Methods for generating large-scale gRNA libraries should be simple, efficient and cost-effective. We describe a protocol for the production of gRNA libraries based on enzymatic digestion of target DNA. This method, CORALINA (comprehensive gRNA library generation through controlled nuclease activity) presents an alternative to costly custom oligonucleotide synthesis.
11:59
Competitive Genomic Screens of Barcoded Yeast Libraries
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Cited by 44 •
2011
We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.
FAQs
Primers define the boundaries of the DNA sequences selected for amplification, so they determine which genes or genomic regions enter the library. This targeted selection concentrates analysis on regions relevant to the research question rather than distributing sequencing across unrelated DNA. In cancer studies, primer design therefore influences which tumor-associated mutations or other genomic alterations can be examined.
PCR increases the amount of each selected target sequence, making the regions available for downstream sequencing. Sequencing adapters are added to support sequencing, while sample-specific indexes identify the source of each library during analysis. Together, these components connect target enrichment with sample tracking, allowing data from multiple cancer specimens or experimental conditions to be distinguished.
A focused design directs sequencing toward selected genes or genomic regions instead of broad, untargeted genomic content. Because the library concentrates on predefined targets, fewer sequencing resources may be needed to address the study question. This efficiency is useful when cancer research emphasizes particular tumor-associated genes and requires comparisons across many samples or treatment conditions.
Preparation begins by selecting target regions and using primers to amplify those sequences from the sample by polymerase chain reaction. The amplified products are then prepared with sequencing adapters, and sample-specific indexes can be added for library identification. These stages create a sequence-ready, traceable collection of selected DNA fragments for subsequent analysis.
They are useful when investigators need to examine targeted genomic changes in tumor-associated genes, including mutations, copy-number changes, and other alterations. The approach is particularly relevant for limited or heterogeneous specimens, where a focused assay can support analysis of selected regions. It also enables comparisons among tumors, treatment conditions, or patient samples.
Sample-specific indexes provide identifying information for individual libraries, allowing samples to remain distinguishable during analysis. This supports multiplexed studies in which libraries from different tumors, patients, or treatment conditions are handled together. After sequencing, the index information helps associate results with the appropriate sample, strengthening comparisons of genomic alterations across the study set.