Table of Contents
When should I use Q5 GC enhancer?
Amplification of some difficult targets, like GC-rich sequences, may be improved by the addition of 1X Q5 High GC Enhancer. The Q5 High GC Enhancer is not a buffer and should not be used alone. It should be added only to reactions with the Q5 Reaction Buffer when other conditions have failed.
How does GC enhancer work?
The 360 GC Enhancer is used for difficult-to-amplify templates, especially for templates with high GC content or GC-repeats. The enhancer can also increase specificity in reactions that generate non-specific products for those templates which have a localized GC-rich sequence.
How fast is Q5 polymerase?
Extension times are generally 20–30 seconds per kb for complex, genomic samples, but can be reduced to 10 seconds per kb for simple templates (plasmid, E. coli, etc.) or complex templates < 1 kb. Extension time can be increased to 40 seconds per kb for cDNA or long, complex templates, if necessary.
What is high-fidelity DNA polymerase?
High-fidelity PCR enzymes are used for applications requiring high accuracy during DNA amplification such as cloning, sequencing or mutagenesis. Thermo Scientific Phusion high-fidelity DNA polymerases were created by fusing a dsDNA-binding domain to a Pyrococcus polymerase–like proofreading enzyme.
What is Q5 polymerase?
Q5® High-Fidelity DNA Polymerase is a high-fidelity, thermostable DNA polymerase with 3´→ 5´ exonuclease activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification.
What is in Q5 buffer?
The 5X Q5 Reaction Buffer contains 2 mM MgCl2 at final (1X) reaction concentrations and is recommended for most routine applications. For GC-rich targets (≥ 65% GC), amplification can be improved by the addition of the 5X Q5 High GC Enhancer.
What does high fidelity mean?
Definition of high fidelity : the reproduction of an effect (such as sound or an image) that is very faithful to the original. Other Words from high fidelity Example Sentences Learn More About high fidelity.
What is low fidelity DNA polymerase?
What are they? Low-fidelity DNA polymerases are recently discovered DNA polymerases that function very inefficiently and inaccurately on undamaged DNA but, unlike replicative polymerases, are able to synthesise DNA past damaged bases.
What is in Q5 reaction buffer?
Shyam deduced that Q5 reaction buffer contains glycerol, which reduces DNA secondary structure, and tetramethylammonium, which increases primer stringency; and Q5 High-GC Enhancer contains DMSO and glycerol to do more of the same.
What is low and high fidelity?
In conclusion. Prototypes are divided into two categories: low and high fidelity. Low fidelity prototypes are used by designers in the early stages of the project to make sure the content is correct. High fidelity prototypes are created at later stages of the design process to test all current hypotheses.
What is the meaning of low fidelity?
Low-fidelity definition The electronic reproduction of sound or images using technology that results in unwanted distortion or imperfections. noun.
Which polymerase has the highest fidelity?
±= 95% confidence Numbers in brackets have limited statistical significance as only 2 mutations were detected after sequencing 441,670 nucleotides. Q5 provides the highest fidelity of all PCR polymerases available!
What is the meaning of low-fidelity?
What is Q5 high GC enhancer and how is it used?
Amplification of some difficult targets, like GC-rich sequences, may be improved by the addition of 1X Q5 High GC Enhancer. The Q5 High GC Enhancer is not a buffer and should not be used alone. It should be added only to reactions with the Q5 Reaction Buffer when other conditions have failed.
What is the purpose of the QIAGEN GC enhancer?
It is used along with GC buffer when the GC content of the primer is high. I speculate the GC enhancer contains DMSO, which interferes with secondary structure formation in GC-rich regions. Unfortunately, Qiagen doesn’t release this information to the public, probably because they spent research dollars in developing a proprietary formula.
Why is Q5 polymerase used with GC buffer?
Q5 Polymerase is an ideal choice for high specificity amplification and provides robust amplification of a wide variety of amplicons, regardless of GC content. It is used along with GC buffer when the GC content of the primer is high. I speculate the GC enhancer contains DMSO, which interferes with secondary structure formation in GC-rich regions.
Can the enhancer be added to the GC reaction buffer?
For particularly difficult or high GC amplicons, the enhancer can be added to the GC Reaction Buffer to improve specificity and/or yield. The enhancer is not a buffer and should not be used alone.