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:
Length of PCR product |
Template DNA |
Template RNA |
50-150 bp |
£200 |
£300 |
151-500 bp |
£150 |
£250 |
501-5000 bp |
£300 |
£400 |
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.
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