Difference between revisions of "Team:Sheffield/Protocols"

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<p>SCIENCE! and other stuff </p>
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<h2>I.Recipes </h2>
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                <p> <b> <u> I.a Agarose gel (50ml of 1% gel)</u> </b> <br>
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1. Weight 0.5 g of agarose <br>
 +
2. Dissolve in 50 ml of 1x TAE buffer <br>
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3. Microwave until completely dissolved <br>
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4. Cool down to ~50<sup>o</sup>C (doesn't burn when you hold it in your hand) <br>
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5. Add 5ul of SybrSafe <br>
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6. Pour the gel into the casting stand <br>
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7. Cover the casting stand with foil to protect it from light (SybrSafe is sensitive to light)<br>
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8. Wait for the gel to solidify <br>
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9. Load the samples (5-10ul of sample + 1-2ul of Loading Dye)<br>
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10. Load 7-10ul of 2-log ladder <br>
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11. Run the gel at 110 V for ~45 min <br>
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12. Use lower gel percentage for larger samples (plasmids etc.) and higher gel percentage for smaller samples (~200-400 bp PCR products)</p>
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<p> <b> <u>I.b. Buffers </u> </b> <br> </p>
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<p> <b>I.b.1. 50X TAE Buffer (1 liter)</b> <br>
 +
 +
1. 242 g Tris-base <br>
 +
2. 18.61 g EDTA <br>
 +
3. 57.1 ml glacial acetic acid <br>
 +
4. Water top up to 1L </p>
 +
 +
<p> <b>I.b.2. 1X TAE Buffer (1 liter)</b> <br>
 +
 +
1. Measure 20 ml of 50X TAE buffer <br>
 +
2. Add 980 ml dH2O <br>
 +
 
 +
</p>
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</div>
 
</div>
 
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Revision as of 21:02, 19 October 2016

A template page

PROTOCOLS

I.Recipes

I.a Agarose gel (50ml of 1% gel)
1. Weight 0.5 g of agarose
2. Dissolve in 50 ml of 1x TAE buffer
3. Microwave until completely dissolved
4. Cool down to ~50oC (doesn't burn when you hold it in your hand)
5. Add 5ul of SybrSafe
6. Pour the gel into the casting stand
7. Cover the casting stand with foil to protect it from light (SybrSafe is sensitive to light)
8. Wait for the gel to solidify
9. Load the samples (5-10ul of sample + 1-2ul of Loading Dye)
10. Load 7-10ul of 2-log ladder
11. Run the gel at 110 V for ~45 min
12. Use lower gel percentage for larger samples (plasmids etc.) and higher gel percentage for smaller samples (~200-400 bp PCR products)

I.b. Buffers

I.b.1. 50X TAE Buffer (1 liter)
1. 242 g Tris-base
2. 18.61 g EDTA
3. 57.1 ml glacial acetic acid
4. Water top up to 1L

I.b.2. 1X TAE Buffer (1 liter)
1. Measure 20 ml of 50X TAE buffer
2. Add 980 ml dH2O