Why can some DNA fragments/genes only be amplified by high-fidelity polymerases and not by Taq?

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Why do I get a PCR band of the wrong size after cloning, though restriction digestion confirms the presence of the gene?

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Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

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How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

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Weak DAPI staining after immunohistochemistry - how to improve?

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Why after performing site directed mutagenesis ,I don't see any colony after transformation?

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For an in-vitro drug release study, what molecular weight cut-off (MWCO) dialysis bag is required for a 117 kDa protein?

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Pranav Pradip Kulkarni

What is size of your DNA fragment?? What is AT/GC %??

Some times size and GC % affect such reactions.

Have you check with repeat template preparation?

Sandeepa Dey

Phusion has way higher processivity and lower elongation time than Taq, so it is overall a more efficient enzyme.

Priyanka Siwach

It happens. size of the DNA fragment to be amplified matters a lot. Phusion is best choice for amplification of large DNA fragments

Amanda O'Donnell

If you are cloning restriction sites onto the ends of your cloned gene then you can't use Taq as it has 5'-3' exonuclease activity, which will chew away the 5' ends of primer overhangs