Talen steps
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This document outlines the 7 steps to design a TALEN sequence to knockout a gene: 1) Design the TALEN sequence, 2) Amplify the sequence via PCR, 3) Assemble the TALEN gene and insert it into an expression vector, 4) Transfect the vector into desired cells, 5) The TALEN pair binds DNA and causes cleavage, 6) A double strand break occurs and non-homologous binding leads to, 7) Mutation of the DNA and knockout of the target gene, resulting in knockout mice.
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Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
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Talen steps
- 1. Design the TALEN sequenceStep 1
- 2. Step 2 Amplify the desired TALEN sequence by PCR
- 3. Step 3 Assemble TALEN gene and insertion into expression vector
- 5. Step 5 TALENS pair binds to DNA and produce specific cleavage
- 6. Step 6 Double strand break of DNA and non homologous binding of DNA
- 7. Step 7 Mutation of DNA leads to the knock out of the gene Knock out mice