"
Team:Cambridge-JIC/Dry Work
From 2014.igem.org
(Difference between revisions)
| Line 6: | Line 6: | ||
<ul> | <ul> | ||
<li>Tip 1</li> | <li>Tip 1</li> | ||
| + | Delete 5'-UTRs (if in doubt). 5'-UTR's affect expression rates, so if we're comparing promoters/inputs, UTR's will make a big difference. So, for the sake of normalisation, get rid of the 5'-UTRs. | ||
<li>Tip 2</li> | <li>Tip 2</li> | ||
| + | If it works, don't change it. Don't go altering spacing in between parts when making constructs! | ||
</ul> | </ul> | ||
| Line 12: | Line 14: | ||
<ul> | <ul> | ||
<li>Tip 1</li> | <li>Tip 1</li> | ||
| + | Make the primers. Don't really worry about secondary structures unless it's obvious. Use a melting temperature of >58 degrees. | ||
<li>Tip 2</li> | <li>Tip 2</li> | ||
| + | Check that they are correct by searching in the plasmid you're PCR'ing off and the destination construct. | ||
</ul> | </ul> | ||
</html> | </html> | ||
Revision as of 14:54, 22 July 2014
| Home | Team | Official Team Profile | TimeLine | Project | Outreach | Marchantia | Informatics | Parts | Modeling | Notebook | Protocols | Safety | Dry Work | Technology | Attributions |  |
Constructs
- Tip 1 Delete 5'-UTRs (if in doubt). 5'-UTR's affect expression rates, so if we're comparing promoters/inputs, UTR's will make a big difference. So, for the sake of normalisation, get rid of the 5'-UTRs.
- Tip 2 If it works, don't change it. Don't go altering spacing in between parts when making constructs!
Primers
- Tip 1 Make the primers. Don't really worry about secondary structures unless it's obvious. Use a melting temperature of >58 degrees.
- Tip 2 Check that they are correct by searching in the plasmid you're PCR'ing off and the destination construct.
