Sanchored PCR: PCR with cDNA coupled to a solid phase

Simplified protocol of solid-phase cDNA libraries for multiple PCR amplification.

Solid-phase PCR in a picowell array for immobilizing and arraying 100,000 PCR products to a microscope slide.

We present a method for performing highly parallel PCR reactions in a picowell array (PWA) simultaneously immobilizing generated PCR products in a covalent and spatially-resolved manner onto a microscope slide via solid-phase PCR (SP-PCR). This so called PWA-SP-PCR was performed in picowell arrays featuring 100,000 wells cm(-2) of 19 pL reaction volumes with a surface-to-volume ratio of 0.2 μm(...

Chimeric cDNA clones: a novel PCR artifact.

During the cloning of a transcript of one member of a closely related gene family, the human 7-crystallin gene family (1), we encountered a novel artifact of the polymerase chain reaction: the formation of chimeric cDNA molecules. Our experimental strategy in cloning the human 7E-crystallin transcript was a common one (2): first strand cDNA synthesis on human lens RNA using AMV reverse transcri...

Enhanced solid phase PCR: mechanisms to increase priming by solid support primers.

Conventional solid phase amplification regimens such as solid phase PCR (SP-PCR), asymmetric SP-PCR, and bridge PCR are mechanistically limited with respect to amplification efficiency and solid support primer involvement. Here we present enhanced solid phase PCR (ESP-PCR) in which solid support primer priming is facilitated by its nesting and high melting temperature (T(m)) relative to the aqu...

Extraction and Purification of DNA from Complex Biological Sample Matrices Using Solid-Phase Microextraction Coupled with Real-Time PCR.

The determination of extremely small quantities of DNA from complex biological sample matrices represents a significant bottleneck in nucleic acid analysis. In this study, polymeric ionic liquid (PIL)-based solid-phase microextraction (SPME) was applied for the extraction and purification of DNA from crude bacterial cell lysate with subsequent quantification by real-time PCR (qPCR) analysis. Us...