Team:Macquarie Australia/Protocols/RD

From 2012.igem.org

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<center><h1>Restriction Digest</h1></center>
<center><h1>Restriction Digest</h1></center>
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<p>We prepared Master Mixes containing to following volumes,</p>
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<p>We prepared Master Mixes containing the following volumes,</p>
<center><table><tr><td><table border="3" cellpadding="4" cellspacing="0">
<center><table><tr><td><table border="3" cellpadding="4" cellspacing="0">
<tr><td colspan="2">EcoR1 + Spe1 Master Mix</td></tr>
<tr><td colspan="2">EcoR1 + Spe1 Master Mix</td></tr>

Latest revision as of 01:24, 26 September 2012



Restriction Digest

We prepared Master Mixes containing the following volumes,

EcoR1 + Spe1 Master Mix
SubstrateVolume (µL)
10x Buffer20
EcoR12.5
Spe12.5
Water75
Total100
Xba1 + Pst Master Mix
SubstrateVolume (µL)
10x Buffer20
Xba12.5
Pst2.5
Water75
Total100
EcoR1 + Pst Master Mix
SubstrateVolume (µL)
10x Buffer20
EcoR12.5
Pst2.5
Water75
Total100

Protocol

  1. Mix 10 µL of the desired DNA/plasmid and 10 µL of the appropriate Master Mix in a PCR tube.
  2. Using a thermocycler, incubate the mixture at 37°C for 30 minutes and then deactivate the enzymes by heating to 80°C for 20 minutes.
  3. Store in a -20°C freezer until DNA required

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