2014. Primer Based Approach for PCR Amplification of High GC Content Gene: Mycobacterium Gene as a Model. Mol Biol Int 2014;2014:937308.Kumar, A., & Kaur, J. (2014). Primer based approach for PCR amplification of high GC content gene: Mycobaterium gene as a model. Molecular Biology ...
2. Melting TemperaturePrimers with similar Tm, preferably between 55°C-60°C are used. For sequences with high GC content, primers with a higher Tm (preferably 75°C-80°C) are recommended. A Tm variation of between 3°-5° C is acceptable for primers used in a pool. 3. Specifici...
DNAsegmentswithveryhighGCcontenthaveproveddifficulttohandleinawiderangeofmolecularanalyses.GC-richregionsmayformrigid,constrainedsecondarystructuresthataredifficultorimpossiblefortheDNApolymerasestoenterunderstandardPCRconditions.6 HighlyGC-richregionscanobstructPCRdependentanalysesinthefollowingsituations:1.StandardPCR...
GC content. Third, primer specificity must be empirically tested with control DNAs of known and unknown methylation states to assess false positive results. To help discriminate methylation states by base-pair mismatches, it is advisable to desig...
Primer Based Approach for PCR Amplification of High GC Content Gene: Mycobacterium Gene as a Model The genome of Mycobacterium is rich in GC content and poses problem in amplification of some genes, especially those rich in the GC content in terminal regions, by standard/routine PCR procedures....
GC-rich PCR DNA templates containing high GC content (>65%) can be difficult to amplify because of the stronger hydrogen bonds between G and C bases. GC-rich sequences can also be involved in secondary structures. Thus, GC-rich s...
One major challenge in the design of highly multiplexed PCR primer sets is the large number of potential primer dimer species that grows quadratically with the number of primers to be designed. Simultaneously, there are exponentially many choices for multiplex primer sequence selection, resulting in...
PCR primers have been traditionally designed by thermodynamic interaction with the desired templates1,2. Primers are designed to increase two respectively significant base sequence specificity and reasonable GC content indicators. The high specificity can prevent mispriming in regions other than the target...
to amplify using conventional PCR techniques due the presence of secondary structures that resist denaturation and prevent efficient primer annealing. Takara Bio offers several DNA polymerases designed to handle such challenging targets, with amplification of templates that contain up to 90% GC content. ...
PCR amplification failure from cDNA libraries or RNA templates, under the optimal conditions is generally attributed to high GC content. Utilization of various additives without thorough analysis of secondary structures of the template as well as primers and subsequent PCR cycle conditions, generally ...