Avoiding cross-contamination during manual qpcr set-up is essential for reliable results, but can be a tedious, time-consuming process. Using an automated liquid handling system prevents cross-contamination and improves efficiency.
Problem: Real-time qPCR enables amplification, detection, and quantification of the target molecule in a single step. Manual qPCR set-up requires significant knowledge and skill, and operators must take precautions to avoid cross-contamination. Despite this, operator error remains the most likely cause of qPCR contamination.
qPCR reaction contamination is a serious problem, as nucleic acid contamination can produce false positive results. There are multiple potential sources of contamination of qPCR reactions, ranging from sample cross-contamination to laboratory equipment and carryover.
Common precautions taken to reduce the risk of cross-contamination include the use of uracil DNA glycosylase (UDG), systematic material decontamination, and maintaining separate workstations for each step of the qPCR process. Unfortunately, these tedious and sometimes time-consuming processes are not guaranteed to prevent cross-contamination, as they do not address a primary cause: accidental contamination during liquid handling.
Answer: Automated liquid handling systems eliminate the risk of contamination by the operator and increase the efficiency of otherwise time-consuming tasks.
There is a very high risk of cross-contamination during sample transfer to the qPCR plate. Even an experienced operator must be extremely careful not to generate aerosol or droplet during the process.
Automated liquid handling systems dramatically reduce the risk of cross-contamination. The epMotion liquid handling system by Eppendorf is an excellent solution for qPCR set-up. The accurate pipetting system eliminates a potential point of human error in the process to ensure reliable results.
Reproducibility is also essential for qPCR assay reliability. The assay must allow for data comparison between samples, plates, and runs. The epMotion automated system ensures high reproducibility, with an intra-plate coefficient of variation never exceeding 1.5%.
For more information, visit www.eppendorf.com/automatedLH