Difference between revisions of "Team:DTU-Denmark/Experiments"
| Line 61: | Line 61: | ||
<ol> | <ol> | ||
<li><p style="font-size:90%">Quickly spin the DNA down in the table centrifuge</p></li> | <li><p style="font-size:90%">Quickly spin the DNA down in the table centrifuge</p></li> | ||
| − | <li><p style="font-size:90%">Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL and for primers it is 100 μM.</p></li> | + | |
| + | <li><p style="font-size:90%">Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL and for primers it is 100 μM.</p></li> | ||
| + | |||
<li><p style="font-size:90%">Genes/Primers from IDT or other DNA provider</p> | <li><p style="font-size:90%">Genes/Primers from IDT or other DNA provider</p> | ||
<ul> | <ul> | ||
<li><p style="font-size:90%">DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is 40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)</p></li> | <li><p style="font-size:90%">DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is 40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)</p></li> | ||
| + | |||
<li><p style="font-size:90%"> In order to calculate the the molar amount of primer, use the <a href="https://nebiocalculator.neb.com/#!/ssdnaamt">NEB calculator</a></p></li> | <li><p style="font-size:90%"> In order to calculate the the molar amount of primer, use the <a href="https://nebiocalculator.neb.com/#!/ssdnaamt">NEB calculator</a></p></li> | ||
</ul> | </ul> | ||
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<li><p style="font-size:90%">Add the calculated amount of EB buffer</p></li> | <li><p style="font-size:90%">Add the calculated amount of EB buffer</p></li> | ||
| + | |||
<li><p style="font-size:90%">Store the resuspended DNA at -20°C </p></li> | <li><p style="font-size:90%">Store the resuspended DNA at -20°C </p></li> | ||
</ol> | </ol> | ||
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| − | + | </div> | |
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</div> | </div> | ||
<button class="collapsible">Open Section 2</button> | <button class="collapsible">Open Section 2</button> | ||
Revision as of 10:14, 14 September 2018
Synthlab protocols
DNA from IDT will typically be delivered in a white flaky substance, which need to be resuspended, before the DNA is ready for use
Materials
Table centrifuge
EB/TE buffer
Genes/Primers from IDT or other DNA provider
Procedure
Quickly spin the DNA down in the table centrifuge
Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL and for primers it is 100 μM.
Genes/Primers from IDT or other DNA provider
DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is 40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)
In order to calculate the the molar amount of primer, use the NEB calculator
Add the calculated amount of EB buffer
Store the resuspended DNA at -20°C
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Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.