LEO PRIME™ is a Taq-based master-mix (2X) designed for unparalleled performance in routine PCR reactions. With a focus on versatility and robustness, LEO PRIME™ stands out with its exceptional features tailored to meet the demands of genotyping, allele specific PCR, low abundance gene testing and multiplexing.
Crudely extracted plant DNA can be directly used for PCR.
PCR Amplifications upto 5KB is supported.
Multiplexing is easier and can detect low copy targets.
Making genotyping simpler and reliable.
High tolerance for many PCR inhibitors/reagents.
Seamless amplification for cDNA 2nd Strand Synthesis.
Making Colony PCR extremely simple, accurate and fast.
Preloaded with green dye for direct gel loading.
One of its notable attributes is its high PCR inhibitor tolerance, making it particularly adept at handling challenging plant DNA samples containing inhibitors such as phenol, ethanol, IPA, humic acid, Trizol, and EDTA. This capability ensures reliable amplification even in the presence of substances known to impede PCR reactions, guaranteeing accurate and reproducible results even with very crude DNA extracts from plants.
LEO PRIME™ is not only resilient in the face of inhibitors but also boasts impressive amplification capabilities, supporting fragment sizes of up to 5kb with validated performance. This allows researchers to amplify a wide range of DNA targets with confidence, expanding the possibilities for downstream applications.
Moreover, LEO PRIME™ ‘s versatility extends to multiplexing, enabling simultaneous amplification of multiple targets within a single reaction.
LEO PRIME™ can also used with super precessions for colony PCR screening, where researchers can directly put the colony in the mastermix for PCR reactions.
LEO PRIME™ Mastermix ensures reliable amplification from cDNA samples, providing researchers with a versatile tool for efficient and accurate gene expression analysis in diverse experimental settings.
Humic acid, a common inhibitor present in soil and environmental samples,can significantly interfere with PCR reactions by inhibiting polymerase activity and binding to DNA. To mitigate its effects, specialized PCR mastermixes with high inhibitor tolerance, like LEO PRIME, are crucial for successful amplification of DNA from such challenging samples.
Template: 10ng of DNA extracted from OKRA plant (CTAB method). Inhibitor added: 5ng, 10ng, 20ng of Humic Acid & control (No inhibitor)
Inference: LEO Prime™ is tolerant to humic acid upto 10ng per reaction and No amplification was observed for competition.
Phenol residues can carry over during nucleic acid extraction, leading to PCR inhibition and compromised results.
Template: 10ng of DNA extracted from OKRA plant (CTAB method); Inhibitor added: 0.5%, 1% and 2% of phenol and control (No inhibitor).
Inference: Consistent amplification was observed for LEO PRIME even at 2% phenol.
EDTA is a chelating agent commonly used in DNA extraction buffers to inhibit DNase activity and preserve DNA integrity. However, EDTA can inhibit PCR by chelating Mg²⁺ ions essential for DNA polymerase activity.
Template: 10ng of DNA extracted from OKRA plant (CTAB method); Inhibitor added: 1mM, 2mM, 3mM EDTA and control (No inhibitor).
Inference:Complete inhibition was observed for competition products even at 1mM EDTA. However, LEO Prime is tolerant ≤2mM EDTA.
To mitigate the inhibitory effects of ethanol and IPA on PCR performance, it’s crucial to ensure thorough and proper purification of DNA samples to remove residual solvents. Additionally, using PCR master mixes specifically formulated with high inhibitor tolerance, such as LEO PRIME, can help overcome potential inhibition and ensure robust PCR amplification even in the presence of these solvents.
Template: 10ng of DNA extracted from OKRA plant (CTAB method). Inhibitor added: 2%, 5%, 10% Ethanl/IPA and control (No inhibitor).
Inference: For both ethanol and IPA, upto 5%v/v good amplification was observed.
Trizol, a popular reagent for RNA extraction, contains guanidine isothiocyanate, which can inhibit PCR reactions if not properly removed during nucleic acid purification.
Template: 10ng of DNA extracted from OKRA plant (CTAB method). Inhibitor added: 0.2%, 0.5%, 0.8% TRIZOL and control (No inhibitor).
Inference: Good tolerance was observed for both mastermixes upto 0.8% Trizol tested.
Long time storage: -20℃, 2 years
Day to day usage: 4℃, One Month
Always prevent the master-mix from primer and nucleases contamination.
Protocol for this product can be found here.
All the products are for research purpose only. Not for human diagnostics.
