Does Anybody has the experience of mate pair sequence? Except the insert size is different from pair end sequence, and what is the benefit with this method?
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
Never tried mate pair myself. Only used PE so far.
Straight from Illumina: http://investor.illumina.com/phoenix...574&highlight=
Check this website that talks of mate pair (MP) libraries: http://seq.molbiol.ru/#i_c_fr
This older thread might also help clarify some things: http://seqanswers.com/forums/showthread.php?t=503
Hope that helps.
-
Illumina's regular "paired end" kits can do an insert size around 500 bp.
Their first "mate pair" kits use a different technique, and can do much longer insert sizes. I understand 4 kbp kits are available, and 10 kbp kits will be available soon.
The BENEFIT of longer insert size comes when trying to assemble sequences. It is IMPOSSIBLE to resolve a repeated sequence of length N unless you have paired end reads with an insert size of at least size N. As many common repeats in genomes are usually much larger than 500 bp, the "mate pair" kits are crucial to resolving these repeats and finishing/closing genomes (or at least drastically reducing the number of contigs).
Comment
-
Originally posted by bair View PostIs it correct that: PE is used for SNP/indel detection and ME for structural variation. Both can be used for de novo assembly?
ANY reads can be used for SNP/indel coverage - that includes short reads, long reads, paired-end reads, single-end reads, mate-pair reads - it doesn't matter, as long as there is sufficient coverage and/or quality to infer a statistically signficant result.
For structural variation, yes of course you need to be able to examine large scale sequence relationships, and PE or SE reads will not do this beyond ~500 bp say. Large insert libraries such as MP and 454FLX are needed here. Or other strategies such as Opgen's "optical mapping" maps.
De novo assembly can use any reads you give it, including SE, PE, and MP. The ability of different software to handle mixes of these data types varies however.
Comment
-
Originally posted by bair View PostIs it correct that: PE is used for SNP/indel detection and ME for structural variation. Both can be used for de novo assembly?
Comment
-
Originally posted by bair View PostIn order to get "robust" SNPs/indel, it might be better to use PE / ME to confirm SNPs by two directions, but I did not see SNPs detection programs doing things like this way. Maybe I'm wrong.
Comment
-
Originally posted by kmcarr View PostMate pair reads are essential (or at least extremely helpful) in de novo projects for orienting contigs into scaffolds. However you shouldn't use mate pair libraries as your source for general coverage. Illumina recommends reads no longer than 36nt when sequencing mate pair libraries; this is to reduce the probability of a read crossing the junction point where the two distant ends of the original DNA fragment were joined during circularization. To generate the bulk of your coverage it is best to use 2x100 (or 2x120) paired end reads. Secondly, the method used to generate mate paired libraries requires a lot of DNA and ends up "throwing away" most of it. This has implications for library diversity.
Comment
Latest Articles
Collapse
-
by seqadmin
The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...-
Channel: Articles
04-22-2024, 07:01 AM -
-
by seqadmin
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...-
Channel: Articles
04-04-2024, 04:25 PM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, 04-25-2024, 11:49 AM
|
0 responses
19 views
0 likes
|
Last Post
by seqadmin
04-25-2024, 11:49 AM
|
||
Started by seqadmin, 04-24-2024, 08:47 AM
|
0 responses
18 views
0 likes
|
Last Post
by seqadmin
04-24-2024, 08:47 AM
|
||
Started by seqadmin, 04-11-2024, 12:08 PM
|
0 responses
62 views
0 likes
|
Last Post
by seqadmin
04-11-2024, 12:08 PM
|
||
Started by seqadmin, 04-10-2024, 10:19 PM
|
0 responses
60 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 10:19 PM
|
Comment