PCR and RT-PCR Optimisation

If you get multiple or spurious bands, smearing or get nothing at all – call us and we will optimise your PCR for you. Do not let poor PCR results slow down your project!

PCR or RT-PCR (Reverse Transcription PCR) from difficult templates can be affected by various factors like buffer content, presence of additives for increased sensitivity or yield, concentration of nucleotides, magnesium ions or choice of polymerase. PCR optimisation may not only increase yield of the reaction, but also greatly improve error rate and specificity.

We deal with a broad spectrum of cases for optimising PCR or RT-PCR:

• Length of PCR products: 50 bp to 15,000 bp
• Difficult templates: high GC- or AT- content, repetitive sequences
• Templates: plasmids, genomic DNAs, total or polyA+ RNAs and cDNA libraries
• Real-Time PCR optimisation is possible upon request
   

Customer provides:
Your requirements on the quality of the results needed and information about the template. For experimental work we will need template DNA/RNA and an aliquote of the oligonucleotides which you used
 

Our service includes:
Design and synthesis of oligonucleotides (if applicable)
Optimisation of reaction mix composition (DNA polymerase, buffer, additives, dNTPs, Mg/Mn) and temperature conditions
Sequencing of target PCR product (if applicable)
Defining composition of reaction master mix and supplying it on a regular basis if needed
 

Duration: 3-21 days

Customer receives:
Report on optimisation, sample of the mix and sample of PCR product
 

Price guide:

NOTE: The price does not include the cost of sequencing of a PCR product longer than 500 bp (£30 per extra 1 kb) or the cost of template DNA or RNA.