Primer design

Design of specific oligo pairs for the PCR amplification of a given DNA region and the optimisation of the amplification.

Plain PCR

In vitro amplification of a given DNA fragment without cloning from any sample from which DNA can be isolated or which we receive as purified DNA.

In our laboratory, there are several applications of the simple PCR technique, e.g.:

• PCR reaction following DNA isolation from a mucosal sample, e.g.:
• Lactose intolerance test
• BRCA-targeted mutation testing: detecting mutations in the BRCA1 and BRCA2 genes using primers designed for the corresponding regions. 
• PCR for sequencing: Pre-sequencing PCR for capillary sequencing of samples
• Multiplex PCR: Amplification of multiple target regions in one reaction
  
  

  qPCR
  
  

  With real-time qPCR (real-time quantitative polymerase chain reaction), the PCR product generated after each PCR cycle can be determined. The amount of PCR product generated during amplification can be determined using a fluorescent dye or probes.

  Specific probes (e.g., TaqMan probe) allow for more detailed detection. During PCR, the software of the real-time qPCR instrument will enable us to monitor PCR amplification in real time. Real-time PCR can be used for qualitative and quantitative analyses; the most suitable application method depends on the user’s needs. The LightCycler® 480 (Roche) real-time PCR instrument is routinely used in our laboratory.

  Capillary sequencing

The nucleotide sequences of samples received as part of the capillary sequencing service are determined using the Sanger method. The pieces may be single DNA isolates, PCR products, or plasmids whose nucleotide sequences are to be determined. The ABI 3500 Genetic Analyzer capillary sequencer is used for this purpose.

Fragment analysis

Following a specific labelling PCR, the ABI 3500 Genetic Analyzer can separate DNA fragments by length at a resolution of up to 1 nt. 

Analysis of formalin-fixed paraffin-embedded samples

DNA is isolated and analysed with the technology of choice.