Team:Washington/HOMEPAGE PROTOCOL

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Cloning Protocols Workflow

Tips and Etiquette:

Keep notebooks up-to-date

Workflow:

Primer Design
Streaking a Plate for Isolation
Overnight Cultures
1) Isolation of Plasmid DNA (miniprep)
2) General PCR Protocol (also see PCR GoTag - product (30-200ng/ ul) Check on gel PCR Purification and/or Dpnl Digest
3) General Digestion Protocol Check on gel PCR Purification or Heat Inactivation (check enzyme temp and time, usually 80C for 20min)
4) Agarose Gel Electrophoresis
5) General Ligation Protocol
6) Heat shock/chemical competent transformation
7) Colony PCR with Green taq
8) DNA Sequencing
9) Making Glycerol Frozen Stocks

I AM PARAGRPAHIRGJIG 2 eam:Washington/GENERAL_BASIC_PROTOCOL">General Basic Protocol Information</a></li>

<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">How to Label EVERYTHING</a></li> </ul> Keep notebooks up-to-date </p>

Workflow:

<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">Primer Design</a>
<a href = "https://2013.igem.org/Team:Washington/HOW_TO_LABEL_EVERYTHING">Streaking a Plate for Isolation</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">Overnight Cultures</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">1) Isolation of Plasmid DNA (miniprep)</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">2) General PCR Protocol </a>(also see <a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">PCR GoTag</a> - product (30-200ng/ ul) Check on gel PCR Purification and/or <a href = "">Dpnl Digest</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">3) General Digestion Protocol</a>Check on gel PCR Purification or Heat Inactivation (check enzyme temp and time, usually 80C for 20min)
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">4) Agarose Gel Electrophoresis</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">5) General Ligation Protocol</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">6) Heat shock/chemical competent transformation</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">7) Colony PCR with Green taq</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">8) DNA Sequencing</a>
<a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">9) Making Glycerol Frozen Stocks</a>

<p1> I AM PARAGRPAHIRGJIG 2 </p1>

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