Overview

So, now that you're hopefully excited to get cloning, let's talk about the flowchart for the process of cloning.

FlowChart:

My talk/this blog will focus on the first three platforms, in silico, molecular and E. coli. 

• In Silico:

• To start anything, you should have a plan of attack
• In silico refers to any computer work you will need to ensure the accuracy of your construct and to develop any primers needed 

• Molecular:

• Once you have your strategy, you will have to spend some time at the lab bench to actually synthesize or obtain any inserts you want to utilize
• And you will need to put those inserts into a plasmid for replication
• That plasmid will then have to be moved to a culture line of bacteria to do that replication

• E. coli:

• So you may have a handful of correctly synthesized plasmid, and you will need to see which of the E. coli uptook the correct plasmid, this involves selection and screening
• Sequencing is also highly highly highly recommended 
• Then you need to move it to Agrobacterium or Biolistics if you want to transform plants

• Agrobacterium:

• Select your Agro and transform it with your plasmid
• Confirm your plasmid made it to the Agro without getting scrambled

• Plant:

• Heavily depends on your plant, it is suggested to follow published protocols for your plant of interest 
  

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Jargon and Concepts

Cloning

1. In Silico

2. Molecular

3. E coli

4. Agrobacterium

5. Plant

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