Amplicon seq and targeted seq
What is the difference between amplicon sequencing and targeted sequencing? How to decide when to use either of them?
sequencing
next-gen
• 395 views
•
link
written
by
mandar.bedse •
0 answers
No answers yet.
Log in to answer this question.
More posts like this
-
HISAT2 or STAR
written by SpliceAndScript •HISAT2 and STAR both are used for aligning our reads to reference genome. But I do not know what the actual difference between them is. …
-
amplicon sequencing or metagenomics?
written by linnet.roque6 •amplicon sequencing or metagenomics? What is the difference between the two main methods for studying the microbiome: amplicon sequencing and metagenomic sequencing. What kind of …
-
Difference between PERMANOVA and DESeq2 analysis for amplicon sequencing
written by BioinfonextHi, I am not able to understand what is the difference between `PERMANOVA` and `DESeq2` analysis for `amplicon microbial analysis`. I do have amplicon sequencing …
-
processing amplicon seq data from miseq
written by sc222 •I have .fastq files from a targeted amplicon seq run on MiSeq using Illumina library prep workflow. How do I process it so that I …
-
Reference genome for WGS and WES.
written by shuksi1984 •What is the difference between Hg38gatkbundle and hg38? Will my output differ, if I use either of these as a reference source for whole exome, …
-
what should be taken care of during the bioinformatic analysis for targeted resequencing, compared …
written by Zhilong Jiawhat should be taken care of during the bioinformatic analysis pipeline for targeted resequencing, compared with whole genome resequencing? Esp. considering the difference of data …
-
panelcn.MOPS description of methodology
written by agata88Hi all! I was reading about the cn.MOPS which can be used for detection of CNVs from genome or exome sequencing. The paper describing this …
-
Exome versus RNA sequencing
written by Abdul Rafay KhanWhat is the difference between Exome sequencing versus RNA sequencing ? What type of de novo assemblers should be use for exome sequencing reads and …
-
Analysis tools to compare two similar Illumina targeted sequencing runs?
written by 14134125465346445What are the commonly used analysis tools to compare two similar targeted sequencing runs done with Illumina? The type of analysis I am thinking could …
-
Shallow Sequencing Vs Deep Sequencing
written by Joe •<p>Hi,rookie question.</p> <p>What is the difference between deep sequencing and shallow sequencing? Is it just the difference of coverage?</p> <p>When do you apply deep/shallow sequencing?</p>
Hello mandar.bedse!
We believe that this post does not fit the main topic of this site.
Not primarily bioinformatics
For this reason we have closed your question. This allows us to keep the site focused on the topics that the community can help with.
If you disagree please tell us why in a reply below, we'll be happy to talk about it.
Cheers!
Could you let me know on why this is irrelevant for this site?
Hi mandar.bedse
This is more related to wet lab sequencing approaches/protocols rather than bioinformatics. You could find the answer if you could google well enough.
Amplicon sequencing is a type of targeted sequencing. You may enrich your data with multiplex PCR-based or hybridization-based techniques. The difference for the user is mainly which type of sequencing machines you currently have - usually people make this choice looking at what the manufacturer is recommending for this particular machine (e.g., IonTorrent machines are usually using multiplex-PCR based approaches, while people tend to use hybridization techniques with Illumina - even if you can exchange the preparation protocol, this is kinda historical thing, as far as I know).
There are various challenges in bioinformatics here too. Multiplex PCR data is usually distributed around log-normal while hybridization-based techniques can be modelled with Negative Binomial quite efficiently. Hybdridization with long probes (120 for Agilent kits) gives you more "stable" enrichment while, when you have even a short indel or mismatch in your amplicon's hybridization site (around 20 basepairs if I remember correctly) - you may have almost 0 product from this region.
I'd say we use amplicons when we need extremely high coverage (>10.000 on average) and hybridization otherwise. Another challenge for the bioinformatics software working with amplicons - deduplication process. Various barcoding techniques are used for that (it is not possible to robustly resolve this issue with bioinformaitcs only)