Gel Electrophoresis and PCR Clean Up
Gel Electrophoresis:
A 1% agarose solution was made and poured into the gel box. 5µL of the 100 base pair DNA ladder was carefully inserted into the first well of the gel. In lane 1, 1µL of loading dye and 5µL of the sample found in PCR tube 1 was carefully added. 1µL of loading dye and 5µL of the sample containing the 515F/806R gene fragments was carefully added to the second well of the gel. Then, 1µL of loading dye and 5µL of the no-DNA control found in tube three was added to the third well. Again, 1µL of loading dye and 5µL of the PCR sample containing 533F/1492R gene fragments were added to well four. 1µl of loading dye and 5µl of the no-DNA control found in sample 5 was added into well 5 while 1µL of loading dye and 5µL of sample 6 containing the NS1/NS2 gene fragments was added to well 6. The gel was run at 100mV for approximately 45 minutes.
PCR Clean Up:
The PCR clean was performed using a PureLink Quick Gel Electrophoresis & PCR Purification Combo Kit created by Invitrogen Life Technologies. A spin column that was provided by the kit was used to filter DNA from the primers and buffers from the PCR reaction.
A 1% agarose solution was made and poured into the gel box. 5µL of the 100 base pair DNA ladder was carefully inserted into the first well of the gel. In lane 1, 1µL of loading dye and 5µL of the sample found in PCR tube 1 was carefully added. 1µL of loading dye and 5µL of the sample containing the 515F/806R gene fragments was carefully added to the second well of the gel. Then, 1µL of loading dye and 5µL of the no-DNA control found in tube three was added to the third well. Again, 1µL of loading dye and 5µL of the PCR sample containing 533F/1492R gene fragments were added to well four. 1µl of loading dye and 5µl of the no-DNA control found in sample 5 was added into well 5 while 1µL of loading dye and 5µL of sample 6 containing the NS1/NS2 gene fragments was added to well 6. The gel was run at 100mV for approximately 45 minutes.
PCR Clean Up:
The PCR clean was performed using a PureLink Quick Gel Electrophoresis & PCR Purification Combo Kit created by Invitrogen Life Technologies. A spin column that was provided by the kit was used to filter DNA from the primers and buffers from the PCR reaction.