Incomplete access to DNA from lysed cells is a common problem in PCR studies. This issue is often caused by inadequate cell lysis and low concentrations of DNA in the sample. Traditionally, cells are lysed with detergents and enzymes (i.e., Proteinase K) followed by a DNA isolation step (e.g., Qiagen's QIAamp DNA Kit). This approach works well for many cell types, although it fails with some cell types, is time consuming, involves many steps and DNA is often lost during the DNA isolation step.
An alternative approach, that gets around these deficiencies, does not use enzymes to lyse cells or involve DNA isolation. Instead it relies on a mild detergent to weaken cell membranes / walls followed by a heating step to lyse cells (e.g., 5x Cell Lysis Buffer by PCRopsis). This lysate is then incubated with PCRopsis Cell tubes to remove PCR inhibitors that interfere with PCR. This approach allows one to thoroughly lyse numerous cell types (i.e., mammalian, bacterial, fungal, etc.) with a common approach in just a few steps, without lost of DNA or use of expensive, specialized consumables (e.g., columns, magnetic beads, etc.). The PCRopsis alternative approach is ideal for samples containing more than one microorganism, where you need to ensure you detect each microorganism.
Below are a few studies using the PCRopsis approach:
Study #1: Detecting bacterial, yeast and human DNA
Study #2: Detecting bacteria from urine patient samples
Study #3: Detecting toe nail fungi
See all studies at: www.pcropsis.com/studies.html