Instructions

Click on the button "Select plant whose gene is to be edited" to display plant.
Click on the button "Select a gene to be edited" .
Click on the button "Retrieve sequence information of gene from NCBI".
Click on the button "Order/synthesize CRISPR plasmid with codon optimised Cas9 gene".
Click on the button "Use softwares like CRISPR-P v2.0 or Cas Offinder to find the optimum Cas9 binding guide RNA locations within the gene".
Click on the button "Synthesize the red highlighted parts of the DNA as guide RNA".
Click on the button "Ligate the synthesized guide RNAs into the CRISPR plasmid".
Click on the button "Transform plant with guide RNA ligated CRISPR plasmid via electroporation or gene gun and regenerate plant".
Click on the button "Isolate genomic DNA from both wildtype plant and transformed plant".
CLick on the button "PCR using gene specific primers from both wildtype plant gDNA and transformed plant gDNA".
Click on the button "Purify the PCR products and analyse them via Sanger DNA Sequencing"
Click on the button "Compare the sanger sequencing results of both PCR products"
Click on the button "If gene edited, identify traits/phenotype due to that non-functional gene"