Investigation into the interactions between graphene oxide (GO) and biomolecules is very important for broad applications of GO in bioassay and bioanalysis. In this work, we describe the interactions between double-stranded DNA (dsDNA) and GO. We demonstrated that dsDNA can bind to GO forming complexes (dsDNA/GO) in the presence of certain salts, which protects dsDNA from being enzymatically digested. On the other hand, we found that a nonionic surfactant, such as triton X-100, can block the formation of dsDNA/GO complexes, so that the enzymatic digestion of dsDNA is restored. These results lead us to believe that the reason for GO protecting dsDNA from enzymatic digestion is the formation of dsDNA/GO complexes hindering the access of DNA enzymes to dsDNA, rather than direct inactivation of the DNA enzymes.

A new gold-nanoparticle (AuNP)-based strategy to dynamically modulate the activity of DNA polymerases and realize a hot-start (HS)-like effect in the polymerase chain reaction (PCR) is reported, which effectively prevents unwanted nonspecific amplification and improves the performance of PCRs. A high-fidelity Pfu DNA polymerase is employed as the model system. Interestingly, AuNPs inactivate the polymerase activity of Pfu at low temperature, thus resembling an antibody-based HS PCR. This inhibition effect of AuNPs is demonstrated for the preamplification polymerization activity of the PCR, which largely suppresses nonspecific amplification at temperatures between 30 and 60 degrees C and leads to highly specific and sensitive PCR amplification with Pfu. Significantly, the fidelity of Pfu is not sacrificed in the presence of AuNPs. Therefore, this AuNP-based HS strategy provides a straightforward and potentially versatile approach to realize high-performance PCR amplification.

Nanoparticle PCR is a novel method to optimize DNA amplification. It performs well in improving specificity, enhancing sensitivity and speed. Several mechanisms were proposed in previous studies: one was based on the interaction between gold nanoparticles (AuNPs) and DNA while the other was attributed to the heat transfer property of AuNPs. In this paper, we propose that the interaction between AuNPs and DNA polymerase can significantly influence PCR. First, the addition of DNA polymerase can eliminate the inhibitory effects of excess AuNPs. Second, the addition of AuNPs will increase yield of the desired PCR product and make the optimum concentration of DNA polymerase move to higher value. Third, while excess polymerase might inhibit amplification efficiency, AuNPs can reverse this process and the yield of PCR amplification. Based on these results we propose a possible mechanism that AuNPs might modulate the activity of polymerase and improve PCR amplification.

Gold nanoparticles (AuNPs) have been proven to be able to improve the specificity or increase the efficiency of a polymerase chain reaction (PCR) when a suitable amount of AuNPs was used. However, there is still a lack of systematic evaluation of AuNPs in real-time PCR. In this study, DNA degradation and the fluorescence quenching effect of AuNPs were first tested in real-time PCR. Then two different kinds of Taq DNA polymerase, native and recombinant Taq polymerase, were employed to evaluate the AuNPs' effect on the threshold cycle (C(T)) values, standard curves and melting curves in real-time PCR. Different ratios of the amount of native Taq DNA polymerase to the amount of AuNPs were also tested. It was found that AuNPs could be applied in real-time PCR with correlation coefficient R(2)>0.989. The combination of 2.09 nM AuNPs with 3.75 U of native Taq DNA polymerase could make the amplification curves shift to the left and enhance the efficiency of the real-time PCR (0.628 39 without AuNPs compared with 0.717 89 with 2.09 nM AuNPs), thus enabling faster detection in comparison with those of control samples. However, no improvement ability of AuNPs was found in real-time PCR based on recombinant rTaq DNA polymerase. Besides, the results suggest that a complex interaction exists between AuNPs and native Taq DNA polymerase.

Polymerase chain reaction (PCR) has become a standard and important molecular biological technique with numerous applications in genetic analysis, forensics and in vitro diagnostics. Since its invention in the 1980s, there has been dramatic performance improvement arising from long-lasting efforts to optimize amplification conditions in both academic studies and commercial applications. More recently, a range of nanometer-sized materials including metal nanoparticles, semiconductor quantum dots, carbon nanomaterials and polymer nanoparticles, have shown unique effects in tuning amplification processes of PCR. It is proposed that these artificial nanomaterials mimic protein components in the natural DNA replication machinery in vivo. These so-called nanomaterials-assisted PCR (nanoPCR) strategies shed new light on powerful PCR with unprecedented sensitivity, selectivity and extension rate. In this review, we aim to summarize recent progress in this direction and discuss possible mechanisms for such performance improvement and potential applications in genetic analysis (particularly gene typing and haplotyping) and diagnostics.

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Nanomaterials, with their diverse dimensions, shapes and surface functional groups, may interact with biomolecules in various ways. In this paper, we reviewed recent advances on the research of the effect of nanomaterials on biomolecular reactions and the use of nano-bio-complexes in biosensors. Much evidence has clearly indicated that nanomaterials are excellent matrixes for the immobilization of enzymes and such complexes often exhibits even higher activities than native enzymes. Consequently, a large amount of highly sensitive biosensors have been developed based on such nano-bio-complexes. We also stressed the importance of nanomaterials on the reactions of nucleic acids. By exploiting the interactions between AuNPs and DNA, several groups have developed a series of novel biosensors for the detection of DNA target and other biologically important molecules. While the state-of-the-art researches are mainly focused on the study of one nanomaterial and one biomolecule, there has been evidence that integration of nano-bio-complexes might lead to much more interesting findings. For example, nanomaterials have been found to exert great impact on complicated systems such as PCR. One might expect that the realization of highly cooperated nano-bio-machines that comprise of different nanomaterials and different biomolecules would lead to highly promising diagnostic tools both in-vivo and in-vitro.

The interaction of colloidal gold with Taq DNA polymerase (Taq) was investigated in this study. Taq-gold conjugate was formed by adding the enzyme to the colloidal gold solution, as evidenced by UV-Vis spectroscopy, X-ray photoelectron spectroscopy, and photon cross correlation spectroscopy measurements. The conjugate was further characterized by transmission electron microscopy. It was found that the Taq-gold conjugate particles were still spherical and well-dispersed. The influence of gold nanoparticles on the bioactivity of Taq was studied by analyzing the yield of the polymerase chain reaction amplification. Results indicated that the enzymatic activity of Taq decreased after interaction with the colloidal gold.