Experiments
Experiments

Experiments in our project were related to either DNA and RNA amplification techniques or expression of our modified Bst DNA polymerase. Methods for conducting each of these experiments can be found in protocols.

Amplification Reactions

  1. E. coli K-12 RNA and genomic DNA extraction
  2. Standard PCR on E. coli K-12 genomic DNA
  3. LAMP on E. coli K-12 RNA and genomic DNA

Protein Expression

  1. Restriction digest of pET24d vector and modified Bst using BamHI and XhoI restriction enzymes
  2. Ligation of Bst insert into pET24d
  3. Heat shock transformation into E. coli chemically competent Top 10 cells
  4. MiniPrep and gel electrophoresis to verify cloning
  5. Heat shock transformation into E. coli BL21 DE3 cells with IPTG for protein expression
  6. Purification of modified Bst and Western blot to verify purification