Team:UFAM Brazil/Protocol7

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Make sure that:</p>
Make sure that:</p>
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<p style="margin-left:100px">• The final concentration of BSA and buffer is 1x.</p>
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<p style="margin-left:50px">• The final concentration of BSA and buffer is 1x.</p>
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<p style="margin-left:100px">• 5 to 10 enzyme’s unit are necessary for 1 µg plasmidial DNA digest.</p>
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<p style="margin-left:50px">• 5 to 10 enzyme’s unit are necessary for 1 µg plasmidial DNA digest.</p>
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<p style="margin-left:100px">• Add to digest system the quantity of DNA enough for 0.5 – 1 µg. If the system will be purified from the agarose gel lately, add quantity of DNA for the digest system of 1 – 2 µg.</p>
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<p style="margin-left:50px">• Add to digest system the quantity of DNA enough for 0.5 – 1 µg. If the system will be purified from the agarose gel lately, add quantity of DNA for the digest system of 1 – 2 µg.</p>
<p>2. Pipet aliquots to each sample tube and add DNA.</p>
<p>2. Pipet aliquots to each sample tube and add DNA.</p>
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<td class="back" ><a href="https://2014.igem.org/Team:UFAM_Brazil/Methods">Methods</a></td>
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<td class="backProtocols" ><a href="https://2014.igem.org/Team:UFAM_Brazil/Methods">Protocols</a></td>
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Latest revision as of 15:24, 16 October 2014

BioBrick Restriction Digest

Reagents:

- dsH2O;

- 10x Buffer (1 – 2 – 3 – 4);

- 10x BSA;

- Restriction Enzymes (EcoRI, Xbal, SpeI, PstI);

- DNA.

Steps:

1. Make a master mix with everything except the DNA. Make sure that:

• The final concentration of BSA and buffer is 1x.

• 5 to 10 enzyme’s unit are necessary for 1 µg plasmidial DNA digest.

• Add to digest system the quantity of DNA enough for 0.5 – 1 µg. If the system will be purified from the agarose gel lately, add quantity of DNA for the digest system of 1 – 2 µg.

2. Pipet aliquots to each sample tube and add DNA.

3. Incubate at 37°C for 2 – 3 hours. Check the eletrophorectic profile.

Protocols

Retrieved from "http://2014.igem.org/Team:UFAM_Brazil/Protocol7"