Question: primers for whole metagenome
0
gravatar for M.Nadeem Khan
8 weeks ago by
Pakistan/Islamabad/Quaid-I-Azam University
M.Nadeem Khan0 wrote:

Hello All I want to do amplify all the bacterail genomic contents present in environmental samples. What kind of primers I have to use? Primers used for 16s are enough for this procedure? or I have to desgin special type of primers?

sequencing genome • 147 views
ADD COMMENTlink modified 2 days ago by Fabio Marroni2.0k • written 8 weeks ago by M.Nadeem Khan0
1

Primers used for 16s are enough for this procedure?

No

ADD REPLYlink written 8 weeks ago by Vijay Lakhujani3.4k
1

This is not a bioinformatics question. Anyway, buy some multiple displacement amplification kit (just be aware of its shortcomings)..

ADD REPLYlink modified 8 weeks ago • written 8 weeks ago by 5heikki7.9k
1

This is just shotgun sequencing right? So no use of PCR

ADD REPLYlink written 8 weeks ago by gb520
1

It's a step you perform prior to any sequencing

ADD REPLYlink written 8 weeks ago by 5heikki7.9k
1
gravatar for Fabio Marroni
2 days ago by
Fabio Marroni2.0k
Italy
Fabio Marroni2.0k wrote:

I am sorry for this shameful self-promotion, but you can give a look at the methods section of our recent paper (actually still waiting for reviewers!) in which the library preparation for WGS is explained.

However, I agree with @gb that being shotgun sequencing by definition you must avoid using primers for amplifying any given region.

ADD COMMENTlink written 2 days ago by Fabio Marroni2.0k
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