Week 2: DNA Gel Art

DNA Gel Art

Protocol Part 0: Designing My Gel Art / Expected Results and Walkthrough

Created a virtual digest in Benchling as a basis for DNA Gel Art (see below)

Benchling Virtual Digest (A Hidden Hello)

Protocol Part 1a: Preparing a 1% Agarose Electrophoresis Gel

Preared a 1% Agarose Electrophoresis Gel (see below)

Protocol Part 1a: Restriction Digest

Ran Restriction Digest (see images below)

Protocol Part 2: Gel Run

Performed Gel Run (see mp4s below)

Protocol Part 3: Imaging My Results With a Transilluminator

Took gel and prepared to image results (see below)

Final Results

Final result (see below)

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Benchling Protocol Notes (sourced from Wiliam & Mary Node TA, Kate Carline)

NotI-HF: rCutSmart, incubates at 37C, 20,000 U/ml = 10 U/ul Kpn1 (Promega): Buffer J, incubates at 37C, 12,000 U/ml = 12 U/ul Sal1 (Promega): Buffer D, incubates at 37C. 10,000 U/ml = 20 U/ul

1.5 ug DNA 324 ng/ul of Kampy B 4.62 ul DNA for N and K 141.4 ng/ul Kampy C (Nanodrop after running out of Kampy B) 10.61 ul for S

15 units of enzyme 1.5 uL Not1-HF 1.25 ul Kpn1 0.75 ul Sal1

2 ul of each 10X Buffer

Remaining to 20 ul NFW 11.88 ul Not1 12.13 ul Kpn1 6.64 ul Sal1

Spin down briefly in picofuge

Incubated for 30 min at 37C

1% agarose gel 2 ul dye with 10 ul reaction 40 min 185 mA 150V //