Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2016/01/16
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Incubate at 37 °C for 10 minutes. Run the reaction on a gel at 100 V for 45 min, then gel purify the ~450 bp fragment and quantify.
Gel Purification & DNA Quantification
Used the Sigma GenElute Gel Extraction kit. Imaging the gel showed a strong band at just smaller than 500 bp (no picture taken). DNA concentration is measured at 29.5 ng/µL, 260/280 is 2.0.
Amounts given are for a 5:1 insert:vector ratio. Can scale back if gel purification yield is low.
Incubate at room temperature for 10 minutes.
Follow the traditional transformation protocol found here. I am leaving the plates on my bench at room temperature (taped) over the weekend.