Connect.BarcodeofLife.net

international online community for dna barcoding professionals

Hi
I am about to start my work on Barcoding the Moths of North-East India and could absolutely use any help that I can get.

The works of Cameron and Whiting, amongst others, have led me to thinking
about the possibility of using additional mt genes, apart from the
conventional COI. The data on nad2, nad3 and nad4 seems convincing.

I would like to know the limitations or pitfalls of using longer gene
sequences for purpose of establishing phylogeny, since I am targeting
the entire COI amplification as opposed to the 5` half of it.

I intend on using an additional nuclear gene, the EF1-alpha and am stuck
at choosing the right primer pairs for it. Ideally a sequence length of
650-700bp should suffice but previous works have sequenced as long as
1280bp for identification. If anyone could clear out my doubts/queries, I
would be grateful.
Regards

Views: 19

Replies to This Discussion

Dear Mansi,
I understand why you wish to target additional genes/ whole mitichondrial genome. everyone in DNA barcoding knows that almost all phylogenetically important regions have already been explored and why COX1 5' region has been ultimately accepted. Now in the form of iBOL and BOLD molecular taxonomy have already moved much further to Build a molecular databse on this COXI region. At this moment the priority is to strengthen this international effort by contributing the required database on different species. Only after this strong directory/ database has been developed, we will come to know about any shortcomings and look for supplementary regions in cases showing significant conflicts in taxonomy/ phylogeny. Now the length of DNA region- any region falling between ~ 500-800 bp can be sequenced in a single pas sequencing and confiirmed by both strand single read sequences to provide appreciable consistency through a standardized procedure. As with other many scientists, I am confident about the COXI region as well as length of this region for phylogenetic importance. shifting to new regions or regions of longer length will definetely lead to confusion and complicacy in fast generating the required database. If you wish to shift to newer regions, it may hold significance in the long run, but may not be of immediate exploitation.
Hi Mansi,

I join Virash's reply as for focusing on the COI barcode region if your project is about building a reference library for species identification, or for getting an insight into cryptic diversity (at least preliminarily). Barcodes proved very informative in leps, and for the kind of purpose I just mentioned it seems like the logical place to start, especially given the current coverage on BOLD for that group.

Other genes will surely prove very insightful and useful if you want to look further into resolving cases of cryptic diversity, or if you want to build a phylogeny - though the geographical focus isn't much justified for the later.

In any case, it is great that you plan on tackling barcoding of NE-Indian leps! Good luck with your project!

Best wishes,
Rodolphe
Hi Rodolphe

I guess yes, it sounds more justified that I start with only the COI barcode region.
The area is just so unexplored that lack of preliminary knowledge about the vast majority of species forced me into being absolutely thorough with the kind of study I want to do on them.
Ultimate aim remains to document the Biodiversity, apart from studying variations in it with altitude etc.
I do intend on incorporating ND5 and wg genes in later stages of my study. But as of now, I am starting with COI barcode region and EF1-alpha.

Thank you

RSS

Translate

Tory's site-wide code

New to the Connect network?


Watch our Intro Webinar


Introduce yourself to the Connect community


Write a blog post


Ask a question

Tory's code

© 2013   Created by Matthew Fisher.   Powered by

Badges  |  Report an Issue  |  Terms of Service