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==Step 5: Page Denaturing Gel== *Prepare a 2D well denaturing gel by scraping off the gel residue outside and peel of the sticker at the back of gel *Load 2 gels into the gel running module *Add in fresh 0.5X TBE buffer to the inside core full the outer core up to the metal line *Pre-run the gel for 30 min at 200V *Dilute sample to no more than 40ng/uL *Dilute TBE-Urea sample buffer (2X) to 1X in each tube. For 2 gels; Mix 0.5 uL Invitrogen 10bp ladder with 4.5 uL dH2O, 5 uL 2x TBE-Urea sample buffer For 4 gels; Mix 0.5 uL Invitrogen 10bp ladder with 9.5 uL ddH2O, 10uL 2X TBE-Urea sample buffer *Denature samples and ladder at 75C for 7 min, then quickly transfer to cooling rack, wait for 5 mins *Flush the wells very well with P1000 & P200 pipette to remove all urea. '''NOTE: DO NOT TOUCH THE TUBE WITH YOUR HANDS IN THE FOLLOWING STEP, GRAB THE CAP''' *Load the samples into the wells, run at 200V for 30 min. *Stain gel with 4uL SYBR Gold in ~100 mL of TBE in clean tray. *Place tray on an orbital shaker for ~5 min.
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