Primer Design for RT-PCR
Primer design is a crucial initial step in any experiment utilizing PCR to target and amplify a known nucleotide sequence of interest. Properly designed primers will increase PCR amplification efficiency as well as isolate the targeted sequence of interest with higher specificity. Many factors that may limit the success of a primer pair can be detected a priori with computational methods. For example, primer dimer detection, amplification of alternative products, stem loop interference, extreme melting temperatures, and genotype-specific variations in the target sequence can all be considered computationally to minimize subsequent PCR failures. The use of computational sequence analysis tools to select the best primer pair from the available candidates will not only reduce experimental rates of failure but also avoid the generation of misleading results arising from the amplification of alternative products.
- Optimal arbitrary primer length for Differential Display(图)
- 2-D Gel Electrophoresis: Constructing 2D-Gel Proteome Reference Maps
- Routine Identity Confirmation of Recombinant Proteins by MALDI-TOF Mass Spectrometry
- Automated Fluorescent DNA Sequencing on the ABI PRISM 377
- SAGE and LongSAGE
- Site-Specific Targeting of DNA Plasmids to Chromosome 19 Using AAV Cis and Trans Sequences
- SNP Genotyping Using Multiplex Single Base Primer Extension Assays
- The AMPS Package for Multiple Protein Sequence Alignment
- Microchip Capillary Electrophoresis Protocol to Evaluate Quality and Quantity of mtDNA Amplified Fragments for DNA Sequencing in
- Analyzing Epigenome Data in Context of Genome Evolution and Human Diseases