Scarab Mistral - Insertion Sequence Detection Kit

The insertion sequence (IS) detection kit tests for the presence of all known E. coli IS elements in any DNA sample of interest.

• IS elements are naturally present in the genomes of E. coli strains commonly used for protein and plasmid production.
• IS element transposition is stimulated by the cell stress response and can lead to IS elements transposing into plasmid DNA and into other regions of the chromosome.

Factors such as the production of foreign proteins or the burden of carrying a high copy plasmid induce the stress response. To eliminate these undesired and potentially disastrous transposition events, all known IS elements have been removed from Clean Genome^® E. coli strains, creating the ideal hosts for the safe and reproducible production of recombinant proteins or plasmid DNAs.

The kit also detects the presence or absence of known recombination hot spots (Rhs) in the E. coli genome.

IS detection kit performance

To demonstrate the kit performance, pBR322 plasmids were prepared in three different E. coli strains (DH10B, K-12 and Clean Genome^®). In addition, a fourth pBR322 plasmid, where the production host was unknown, was purchased from a well-known commercial supplier. PCR analysis of the plasmid DNAs shows that IS contamination is routine unless you use Clean Genome^® E. coli. Note that IS elements are routinely detected in commercial plasmid preparations:

Positive Control Genomic DNA: 170 μL, sufficient for the analysis of 10 samples.
Negative Control Genomic DNA: 170 μL, sufficient for the analysis of 10 samples.
IS-specific Forward (F) and Reverse (R) Primers: 80 μL of each primer at a concentration of 5 μM, sufficient for the analysis of 10 samples.

Forward Primers

• IS1 forward primer
• IS2 forward primer
• IS3/ISEc17 forward primer
• IS4 forward primer
• IS5 forward primer
• IS10 forward primer
• IS30D forward primer
• IS150 forward primer
• IS186 forward primer
• IS600/ISsd1 forward primer
• IS609 forward primer
• IS911 forward primer
• ISEc1/3/5 forward primer
• ISEc4 forward primer
• RhsA/B/C forward primer
• RhsD/E forward primer

Reverse Primers

• IS1 reverse primer
• IS2 reverse primer
• IS3/ISEc17 reverse primer
• IS4 reverse primer
• IS5 reverse primer
• IS10 reverse primer
• IS30D reverse primer
• IS150 reverse primer
• IS186 reverse primer
• IS600/ISsd1 reverse primer
• IS609 reverse primer
• IS911 reverse primer
• ISEc1/3/5 reverse primer
• ISEc4 reverse primer
• RhsA/B/C reverse primer
• RhsD/E reverse primer