Difference between revisions of "20th Discussion-6 Feb 2018"

From BioWiki
Jump to navigationJump to search
Line 46: Line 46:
 
* Which tools have you found useful thus far for analysis?
 
* Which tools have you found useful thus far for analysis?
 
* Any particular quality control approaches you are using?
 
* Any particular quality control approaches you are using?
 +
 +
Nanopore
 +
* Who is doing it and what are your experiences?
 +
* Now they have a direct RNA sequencing kit
 +
* minoTour - realtime nanopore monitoring
 +
* Mark, UW Madison - quality of flowcells drops considerally, they run 32-36 hours
 +
* Nanopore is operator / library sensitive
 +
* air bubbles in flowcell can happen
 +
* EDTA will kill it
 +
* Minknow is the software from oxford nanopore, mac/linux/windows, downstream albacore
 +
* smaller flowcells are coming - running a smidgeon on a minion
 +
* minion_qc on github useful QC plots, minimap2 for alignment
 +
* epitome from ONT
 +
* WIMP - what's in my pot
 +
* how about base modifications?
 +
** nanopolish can report methylated bases, but some question about accuracy. Paper in nature methods
 +
** Simon mentioned trying to use it to assemble immunoglobulin, found nanopolish was over collapsing their reads
  
 
== Other New Protocols or Approaches? ==
 
== Other New Protocols or Approaches? ==
 
* Anything new happening at your institution that you would like to discuss?
 
* Anything new happening at your institution that you would like to discuss?

Revision as of 09:26, 6 February 2018

Attendees

Training

  • How much does your core do, and how do you schedule it?
  • What do you teach?
  • Do you have any requirements to entry?
  • What are your favorite platforms for teaching or teaching tips?

How much training do you do?

  • 2-3 full days modeled on software carpentry, done a few times, command line, RNA-seq
  • Simon Andrews, Babraham Institute
    • huge amount of training, 50 courses a year, a day each
    • one of their core functions
    • internal / external / commercial venture
    • 5 regular teachers
  • Matt Eldridge, Cambridge
    • Also a large amount of training
    • Part of a larger university program
    • demand is massive, long waiting list
    • perhaps have noticed routine analysis going down in volume
      • maybe that's fine, let routine stuff go and focus on harder analyses
  • How much? free?
    • 50 pounds a day for postdocs, Phd students free
    • 178 pounds a day with a "day rate"

Curriculum?

  • command line, common bioinfo tools
  • RNA-seq / ChIP-seq / application specific
  • someone suggested github / docker
  • Simon said they do
    • statistics
    • core skills - unix, perl/python, R
    • RNA-seq / chIP-seq / methyl-seq
    • "creating figures"
    • gene set enrichment analysis
  • Goblet was mentioned
    • but direct reuse is harder than it seems
  • Environments are tricky
    • how do you ensure the same environment for everyone?
    • amazon - but then dependent on network
    • some people use docker - last part of course is getting things running on *your* laptop
    • Use a script to check that everything is installed and ready to go

Nanopore

  • How is your core supporting Nanopore sequencing?
  • Which tools have you found useful thus far for analysis?
  • Any particular quality control approaches you are using?

Nanopore

  • Who is doing it and what are your experiences?
  • Now they have a direct RNA sequencing kit
  • minoTour - realtime nanopore monitoring
  • Mark, UW Madison - quality of flowcells drops considerally, they run 32-36 hours
  • Nanopore is operator / library sensitive
  • air bubbles in flowcell can happen
  • EDTA will kill it
  • Minknow is the software from oxford nanopore, mac/linux/windows, downstream albacore
  • smaller flowcells are coming - running a smidgeon on a minion
  • minion_qc on github useful QC plots, minimap2 for alignment
  • epitome from ONT
  • WIMP - what's in my pot
  • how about base modifications?
    • nanopolish can report methylated bases, but some question about accuracy. Paper in nature methods
    • Simon mentioned trying to use it to assemble immunoglobulin, found nanopolish was over collapsing their reads

Other New Protocols or Approaches?

  • Anything new happening at your institution that you would like to discuss?