Talk:Synthetic Biology:Vectors: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
 
(2 intermediate revisions by the same user not shown)
Line 1: Line 1:
==Vector design review meeting==
==Vector design review meeting==


We will be holding a meeting to review the proposed design for some new BioBricks standard vectors.
We held a meeting to review the proposed design for some new BioBricks standard vectors.  


===Scheduling===
===Meeting notes===
 
<font color="green">The meeting will be on '''Friday March 17 from 3:00-4:30pm in 68-474.'''
Preliminary cost estimates from Blue Heron will be available.  Apologies to all those who can't make it and wanted to attend!</font> 
 
===Issues to be discussed===
*Are there any other features that we want in these vectors?
*Are there any other features that we want in these vectors?
**All BioBricks compatible restriction sites should be removed from the vectors once the resistance marker and replication origin have been inserted.
**Only include a single antibiotic resistance marker (rather than two).
**Include a unique restriction site in the backbone for situations where the insert is equal in size to the backbone (such that they can be distinguished).
**Since all components will be synthesized, remove unnecessary restriction sites.  Also do codon optimization?  See [[Codon pairs]].
*Should the flanking terminators be inside or outside the verification primer binding sites?
**Inside.
*[[Synthetic Biology:Vectors/Barcode | Plasmid barcode]]
*[[Synthetic Biology:Vectors/Barcode | Plasmid barcode]]
**How can we get this to a final stage?
**Use a 3-4 bp tag that can be read during sequencing.  Also place this tag immediately 3' to VF2 and VR so that vector specific primers can be used in single colony PCR?
**Does this require a more significant investment of resources?
**Is there an interim-term solution?
*How can we "verify" the design?
*How can we "verify" the design?
**Caitlin, Barry and TK have volunteered to inspect the design.
*How should the vectors be fabricated?
*How should the vectors be fabricated?
**Direct synthesis of complete vectors
**Direct synthesis of useful vector components, scaffold and intact vectors (budget of $20,000).  Prepare a flow chart of synthesis and cloning steps so that we can obtain useful intermediates and test whether they work or not.
***expensive
***synthesis problems are dealt with by company not us
**Direct synthesis of necessary parts with manual assembly
***less expensive
***very time-consuming and labor intensive
***risks not getting done
**Direct synthesis of complete vectors and individual parts
***very expensive (how much more?)
***enables construction of new variants later
**Direct synthesis of one variant with built-in capability to swap in other origins and resistance cassettes.
***less expensive
***requires some time and labor to obtain other variants
***risks not getting done
***enables construction of new variants later
*Others?


===Relevant pages===
===Relevant pages===
Line 37: Line 22:
*[[Synthetic Biology:Vectors/Single copy plasmid | Single copy plasmid]]
*[[Synthetic Biology:Vectors/Single copy plasmid | Single copy plasmid]]
*[[Synthetic Biology:Vectors/Barcode]]
*[[Synthetic Biology:Vectors/Barcode]]
*[[Codon pairs]]

Latest revision as of 09:56, 31 May 2006

Vector design review meeting

We held a meeting to review the proposed design for some new BioBricks standard vectors.

Meeting notes

  • Are there any other features that we want in these vectors?
    • All BioBricks compatible restriction sites should be removed from the vectors once the resistance marker and replication origin have been inserted.
    • Only include a single antibiotic resistance marker (rather than two).
    • Include a unique restriction site in the backbone for situations where the insert is equal in size to the backbone (such that they can be distinguished).
    • Since all components will be synthesized, remove unnecessary restriction sites. Also do codon optimization? See Codon pairs.
  • Should the flanking terminators be inside or outside the verification primer binding sites?
    • Inside.
  • Plasmid barcode
    • Use a 3-4 bp tag that can be read during sequencing. Also place this tag immediately 3' to VF2 and VR so that vector specific primers can be used in single colony PCR?
  • How can we "verify" the design?
    • Caitlin, Barry and TK have volunteered to inspect the design.
  • How should the vectors be fabricated?
    • Direct synthesis of useful vector components, scaffold and intact vectors (budget of $20,000). Prepare a flow chart of synthesis and cloning steps so that we can obtain useful intermediates and test whether they work or not.

Relevant pages