Strategies of genetic engineering - How to get gene out of organism

4 important questions on Strategies of genetic engineering - How to get gene out of organism

What do you perform when you have a known DNA sequence and available template mRNA?

Reverse transcription into cDNA and PCR.

When do you perform a PCR with degenerated primers?

When you have a known protein sequence and available template DNA (but DNA sequence not known).

What is a degenerate primer?

Mixture of primers that has substitution of different bases sequence (they are similar not same).
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What is Golden Gate Assembly?

Seamless assembly of DNA fragments.
Activities of a single type IIS restriction enzyme and T4 DNA ligase.
Uses the ability of Type IIS restriction endonucleases to cleave DNA outside of the recognition sequence.

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