Sheep ear tissue samples were treated with 180 µl Buffer ATL and 20 µl QIAGEN Proteinase K at 56°C overnight. Genomic DNA was purified from the lysed tissue samples using the BioSprint 15 DNA Blood Kit with the BioSprint 15 DNA Tissue protocol. DNA was eluted in 200 µl elution buffer. Eluates (5 µl) were run on a 0.8% agarose gel in 1x TBE. M: markers (Lambda HindIII).
Human blood was collected and treated with one of 3 anticoagulants: citrate (C), EDTA (E), or heparin (H). DNA was purified from 200 µl blood immediately after blood collection (Fresh) and after one cycle of freezing and thawing (Frozen). DNA was eluted in 200 µl elution buffer. Eluates (1 µl and 10 µl) were subjected to PCR (50 µl reaction volume) using primers to amplify a 1 kb fragment of the Hugl gene. PCR products were run on a 1.5% agarose gel in 1x TBE. M: markers.
The BioSprint instrument transfers magnetic particles from one well or tube to another.