PCR amplification at the indicated Mg2+ concentrations using QIAGEN PCR Buffer and Taq DNA Polymerase (QIAGEN). The same PCR was performed in parallel using a PCR buffer and Taq polymerase from another supplier (Supplier AII). The single-copy human prion protein gene was amplified successfully in each case using the buffer and enzyme from QIAGEN. M: markers.
Two different primer-template systems were amplified in duplicate using QIAGEN PCR Buffer and Taq DNA Polymerase in the absence (–) or presence (+) of 1x Q-Solution. Q-Solution enables specific amplification of difficult templates. [A] human angiotensin receptor II gene; [B] mouse protein kinase C gene; M: markers.
The human single-copy cystic fibrosis gene was amplified with Taq DNA Polymerase and QIAGEN PCR Buffer using the indicated annealing temperatures. Primers employed were a 22mer with a Tm of 57.5°C (GC content: 54.5%) and a 32mer with a Tm of 85.2°C (GC content: 78%). For analysis, 10% of a 100 µl reaction was loaded. M: markers.
A fragment of the single-copy gene for cystic fibrosis was amplified from 30 ng, 3 ng, and 300 pg human genomic DNA corresponding to 104, 103, and 102 copies of target template, respectively. Three different lots of Taq DNA Polymerase were used and equal volumes of the PCR product were analyzed on a 1% agarose gel. M: markers.
Three different-sized products from human genomic DNA were amplified using either Taq DNA Polymerase and QIAGEN PCR Buffer (QIAGEN), or Taq polymerase and a buffer from another supplier (Supplier AII). For analysis, 10% of each reaction was loaded on the gel. Results from duplicate PCR amplifications are shown. M: markers.