Dynamic Passivation with BSA Overcomes LTCC Mediated Inhibition of PCR
نویسندگان
چکیده
The increasing use of low temperature co-fired ceramic (LTCC) for the fabrication of biological microfluidic devices necessitates further research on LTCC biocompatibility. In this study we explore the inhibitory effect of DuPont’s 951 LTCC on Polymerase Chain Reaction (PCR), and demonstrate a novel mechanism to increase biocompatibility between LTCC and PCR with the addition of a common passivation substance, bovine serum albumin (BSA). We show that DuPont’s 951 LTCC binds negatively charged proteins including BSA and ovalbumin (OVA). This is a significant discovery as proteins (enzymes) are an essential component of most biological reactions, and a frequent addition to microfluidic devices. A proposed model for LTCC inhibition of PCR by enzyme adsorption is presented.
منابع مشابه
Bovine serum albumin reverses inhibition of RT-PCR by melanin.
Melanin contained in pigment cells in a variety of tissues co-purifies with nucleic acids in standard RNA (or DNA) extraction procedures. The presence of melanin in RNA (or DNA) templates can inhibit reverse transcription polymerase chain reactions (RT-PCR) and/or PCR (5). In the past, to circumvent this problem, melanin has been separated from DNA by either columnchromatography (5) or acid-pre...
متن کاملIncreased amplification efficiency of microchip-based PCR by dynamic surface passivation.
Surface passivation is critical for effective PCR using silicon-glass chips. We tested a dynamic polymer-based surface passivation method. Polyethylene glycol 8000 (PEG 8000) or polyvinylpyrrolidone 40 (PVP-40) applied at 0.75% (w/v) in the reaction mixture produced significant surface passivation effects using either native or SiO2-precoated silicon-glass chips. PCR amplification was achieved ...
متن کاملMinimizing the surface effect of PDMS–glass microchip on polymerase chain reaction by dynamic polymer passivation
Polydimethyl siloxane (PDMS)–glass microchip has a very strong surface effect on polymerase chain reaction (PCR), leading to a very poor PCR yield. In the work reported here, practical dynamic passivation of surfaces of PDMS–glass microchip using polyethylene glycol (PEG) or polyvinylpyrrolidone (PVP) was achieved using a conventional thermocycler. The passivation procedure was cost-effective a...
متن کاملMolecular basis for zinc transporter 1 action as an endogenous inhibitor of L-type calcium channels.
The L-type calcium channel (LTCC) has a variety of physiological roles that are critical for the proper function of many cell types and organs. Recently, a member of the zinc-regulating family of proteins, ZnT-1, was recognized as an endogenous inhibitor of the LTCC, but its mechanism of action has not been elucidated. In the present study, using two-electrode voltage clamp recordings in Xenopu...
متن کاملmiR-223 contributes to the AGE-promoted apoptosis via down-regulating insulin-like growth factor 1 receptor in osteoblasts
Advanced glycation end products (AGEs) have been confirmed to induce bone quality deterioration in diabetes mellitus (DM), and to associate with abnormal expression of miRNAs in DM patients or in vitro Recently, miRNAs have been recognized to mediate the onset or progression of DM. In the present study, we investigated the regulation on miR-223 level by AGE-BSA treatment in osteoblast-like MC3T...
متن کامل