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...

The use of PCR based fingerprinting using short primers is very sensitive to variation in procedures. Strict laboratory procedures, exceeding general molecular biology standards, must be followed. Many laboratories have experienced difficulties in obtaining reproducible banding. Using a stock solution greatly minimizes pipetting small quantities, but the stability of a Taq stock is not well kno...

Structural alterations of the apolipoprotein E (apo E) are known to influence the lipid metabolism. The three most common isoproteins Arg 158) are encoded by three co-dominant alleles (e2, el, e4). Phenotyping is cumbersome and, therefore, several methods for apo E genotyping have been developed. Of these, PCR-RFLP (restriction fragment length polymorphism) is currently most rapid and cost-effe...

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...

................................................................................................................................... iii Útdráttur ................................................................................................................................... v Acknowledgement ........................................................................................................

20 ll volume containing: 20 ng template genomic DNA (prepared from peripheral blood lymphocytes by standard protocols), 1 ́ Taq buffer (10 mM Tris±HCl, pH 8.3, 1.5 mM MgCl2, and 0.001% gelatine), 150 lM each dATP, dGTP, dCTP and dTTP (Amersham, Little Chalfont, UK 1 ), 4 ng forward primer (5¢ end-labelled with [P]ATP (Amersham) using T4 polynucleotide kinase (Amersham)), 4 ng unlabelled reverse ...

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...