Biochemical analysis and optimization of inhibition and adsorption phenomena in glass–silicon PCR-chips

Dynamic passivation with BSA overcomes LTCC mediated inhibition of PCR

“…We suggest that further research be performed on the biocompatibility of DuPont's 951 LTCC from various lots. Other materials, such as silicon, which are being used for the construction of PCR devices, also show partial inhibition of PCR reactions through the adsorption of the enzyme Taq polymerase [19][20][21][22].…”

“…Increasing Taq polymerase levels in PCR reactions has been shown to reduce inhibition by silicon [19]. We investigated this approach in experiments with increasing concentrations of the enzyme Taq polymerase in PCR reactions.…”

“…However, it has been shown that not all LTCC is compatible with standard PCR reactions. When PCR is carried out in the presence of specific types of LTCC the reaction can be inhibited [14], which is also a problem found with silicon PCR devices [19][20][21][22]. This has been a major challenge in fabricating reliable PCR devices from LTCC and silicon.…”

“…It has been shown that the enzyme Taq polymerase adsorbs to bare silicon, which is the main cause of PCR inhibition by silicon [19][20][21][22][23]. Studies have also shown that serum proteins bind to certain ceramic substances such as synthetic hydroxyapatite, and Al 2 O 3 .…”

“…Dynamic passivation involves including passivation reagents in the PCR reaction mixture to make the material compatible with the reaction. Common passivation substances in use today are polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), and bovine serum albumin (BSA), which are assumed to preferentially bind to the surface of the material, leaving essential PCR components free in the reaction mixture [1,4,19]. BSA is a PCR additive that is used when inhibitors may be present in the reaction [27][28][29] and is commonly used passivation reagent in PCR devices [2,4,11,30].…”