BACKGROUND: Clinical specimens are routinely fixed in 10% buffered formalin and paraffin embedded. Although DNA is commonly extracted from fixed tissues and amplified by PCR, the effects of formalin fixation are relatively unknown. Formalin fixation is known to impair PCR, presumably through damage that blocks polymerase elongation, but an insidious possibility is error prone translesion synthe...

Given the scale of ongoing COVID-19 pandemic, need for reliable, scalable testing, and likelihood reagent shortages, especially in resource-poor settings, we have developed an RT-qPCR assay that relies on alternative to conventional viral reverse transcriptases, a thermostable transcriptase/DNA polymerase (RTX) (Ellefson et al., 2016). Here show RTX performs comparably other assays sanctioned b...

Decent hot-start effects were here reported in Taq DNA polymerase-based polymerase chain reaction (PCR) when water-soluble CdTe quantum dots (QDs) were employed. The hot-start effects were revealed by the higher amplicon yields and distinguished suppression of nonspecific amplification after pre-incubation of PCR mix with quantum dots between 30 ̊C and 56 ̊C. DNA targets were well amplified even ...

Title of Document: RESIDUAL DNA IN COMMERCIAL TAQ DNA POLYMERASE AS A SOURCE OF INTERFERENCE WITH IMMUNO-PCR ASSAY Jake Juyoung Guag, MPH, 2012 Directed By: Professor and Director, Dr. Donald K. Milton, Maryland Institute for Applied Environmental Health Polymerase Chain Reaction (PCR) was developed for a broad range of purposes. As part of developing a very sensitive Immuno-quantitative PCR (i...

optimization of the condition for pcr-directed sequencing of microsatellites poly adenine (a) length polymorphisms is more difficult and sensitive compared with other common sequences. replication slippage may occur for polymerase enzyme during microsatellite amplification and direct sequencing of these pcr products will be challenging for heterozygote samples. so, the aim of this study is to i...

TA cloning is one of the simplest and most efficient methods for the cloning of PCR products. The procedure exploits the terminal transferase activity of certain thermophilic DNA polymerases, including Thermus aquaticus (Taq) polymerase. Taq polymerase has non-template dependent activity which preferentially adds a single adenosine to the 3'-ends of a double stranded DNA molecule, and thus most...

It is generally believed that sequence heterogeneity in PCR products from fossil remains are due to regular DNA polymerase errors as well as miscoding lesions compounded by damage in the template DNA (Pääbo 1990; Handt et al. 1994b, 1996; Höss et al. 1996; Krings et al. 1997). However, it has been difficult to test the frequency with which this assumption holds. First, DNA extractions from foss...

The highly thermostable DNA polymerase from Thermus aquaticus (Taq) is ideal for both manual and automated DNA sequencing because it is fast, highly processive, has little or no 3'-exonuclease activity, and is active over a broad range of temperatures. Sequencing protocols are presented that produce readable extension products greater than 1000 bases having uniform band intensities. A combinati...

Potent PCR inhibitors in blood and soil samples can cause false negative results from PCR-based clinical and forensic tests. We show that the effect of these inhibitors is primarily upon Taq DNA polymerase, since mutational alteration of the polymerase can overcome the inhibition to the extent that no DNA purification is now required. An N-terminal deletion (Klentaq1) is some 10-100-fold inhibi...