However, bacterial cells can be more difficult to lyse. Therefore, it might be necessary to incubate the bacterial cells for 10 min at 65°C rather than at room temperature in the lysis buffer D2, to make the lysis more stringent. All succeeding steps should be done according to the standard protocol. The most critical factor to get this protocol to work is the quality of the bacterial DNA after lysis. It is crucial to start with fresh, top quality material.
Our customers already gave us some feedback that the DNA of gram+ as well as gram- bacteria was successfully amplified with the Repli-g Advanced DNA Single Cell Kit using the more stringent lysis temperature.
The Repli-G Advanced DNA Single Cell Kit is a perfect choice especially for small amounts of bacterial starting material because the reagents are decontaminated for any residual DNA, reducing possible artefacts.