(072.00, 739.61) (535.26, 739.61) (535.26, 752.34) (072.00, 752.34)       /TT0 MOHAVE COMMUNITY COLLEGE                                                                                           BIO181 	<|special_separator|>
(296.76, 048.53) (318.18, 048.53) (318.18, 058.91) (296.76, 058.91)       /TT1 133 	<|special_separator|>
(072.00, 035.81) (075.07, 035.81) (075.07, 046.19) (072.00, 046.19)       /TT1  	<|special_separator|>
(529.92, 589.98) (532.35, 589.98) (532.35, 603.16) (529.92, 603.16)       /TT2  	<|special_separator|>
(072.00, 567.03) (283.59, 567.03) (283.59, 578.92) (072.00, 578.92)       /TT2 3. Mix reagents by pipetting gently up and down.  	<|special_separator|>
(072.00, 541.35) (292.47, 541.35) (292.47, 553.25) (072.00, 553.25)       /TT2 4. Incubate all of the reaction tubes for 1 hour at 37 	<|special_separator|>
(292.44, 545.70) (295.88, 545.70) (295.88, 553.44) (292.44, 553.44)       /TT2 o	<|special_separator|>
(295.92, 541.35) (308.07, 541.35) (308.07, 553.24) (295.92, 553.24)       /TT2 C.  	<|special_separator|>
(072.00, 515.67) (392.55, 515.67) (392.55, 527.57) (072.00, 527.57)       /TT2 NOTE: Your instructor will freeze your completed restriction digests at -20 	<|special_separator|>
(392.52, 520.02) (395.96, 520.02) (395.96, 527.76) (392.52, 527.76)       /TT2 o	<|special_separator|>
(396.00, 515.67) (512.91, 515.67) (512.91, 527.56) (396.00, 527.56)       /TT2 C until the next lab period.  	<|special_separator|>
(072.00, 490.91) (219.84, 490.91) (219.84, 501.20) (072.00, 501.20)       /TT3 III. Electrophorese Digests 	<|special_separator|>
(072.00, 463.25) (113.46, 463.25) (113.46, 474.00) (072.00, 474.00)       /TT2 Reagents:  	<|special_separator|>
(090.00, 441.66) (094.14, 441.66) (094.14, 449.34) (090.00, 449.34)      /C2_0 •	<|special_separator|>
(094.08, 441.03) (096.58, 441.03) (096.58, 449.49) (094.08, 449.49)       /TT1  	<|special_separator|>
(108.00, 438.30) (254.82, 438.30) (254.82, 449.05) (108.00, 449.05)       /TT2 Restriction digests from Part II, on ice 	<|special_separator|>
(090.00, 430.62) (094.14, 430.62) (094.14, 438.30) (090.00, 438.30)      /C2_0 •	<|special_separator|>
(094.08, 429.99) (096.58, 429.99) (096.58, 438.44) (094.08, 438.44)       /TT1  	<|special_separator|>
(108.00, 427.25) (184.63, 427.25) (184.63, 438.00) (108.00, 438.00)       /TT2 10x loading dye, 10 	<|special_separator|>
(184.56, 427.25) (189.50, 427.25) (189.50, 438.00) (184.56, 438.00)      /C2_1 𝜇𝜇	<|special_separator|>
(189.72, 427.25) (196.50, 427.25) (196.50, 438.00) (189.72, 438.00)       /TT2 L 	<|special_separator|>
(072.00, 402.66) (167.58, 402.66) (167.58, 413.41) (072.00, 413.41)       /TT2 Supplies and Equipment 	<|special_separator|>
(090.00, 380.94) (094.14, 380.94) (094.14, 388.62) (090.00, 388.62)      /C2_0 •	<|special_separator|>
(094.08, 380.31) (096.58, 380.31) (096.58, 388.76) (094.08, 388.76)       /TT1  	<|special_separator|>
(108.00, 377.57) (381.66, 377.57) (381.66, 388.32) (108.00, 388.32)       /TT2 Gel electrophoresis chamber with agarose gel in gel tray, power supply 	<|special_separator|>
(090.00, 369.90) (094.14, 369.90) (094.14, 377.58) (090.00, 377.58)      /C2_0 •	<|special_separator|>
(094.08, 369.26) (096.58, 369.26) (096.58, 377.72) (094.08, 377.72)       /TT1  	<|special_separator|>
(108.00, 366.53) (127.98, 366.53) (127.98, 377.28) (108.00, 377.28)       /TT2 1-20 	<|special_separator|>
(127.92, 366.53) (132.85, 366.53) (132.85, 377.28) (127.92, 377.28)      /C2_1 𝜇𝜇	<|special_separator|>
(132.84, 366.53) (139.62, 366.53) (139.62, 377.28) (132.84, 377.28)       /TT4 L 	<|special_separator|>
(139.68, 366.53) (246.06, 366.53) (246.06, 377.28) (139.68, 377.28)       /TT2 Micropipette and pipet tips 	<|special_separator|>
(072.00, 342.78) (136.83, 342.78) (136.83, 352.24) (072.00, 352.24)       /TT3 Load the Gel 	<|special_separator|>
(072.00, 313.71) (201.14, 313.71) (201.14, 325.60) (072.00, 325.60)       /TT2 1. Use a micropipette to add 2 	<|special_separator|>
(201.11, 313.71) (206.57, 313.71) (206.57, 325.60) (201.11, 325.60)      /C2_1 𝜇𝜇	<|special_separator|>
(206.64, 313.71) (211.99, 313.71) (211.99, 325.60) (206.64, 325.60)       /TT4 L	<|special_separator|>
(211.92, 313.71) (530.77, 313.71) (530.77, 325.60) (211.92, 325.60)       /TT2  of 10× loading dye to a reaction tube. Use the pipet tip and gently pipet up 	<|special_separator|>
(072.00, 301.95) (537.88, 301.95) (537.88, 313.84) (072.00, 313.84)       /TT2 and down a couple of times to mix the 10× loading dye with the digested DNA. Use a new pipet tip and repeat 	<|special_separator|>
(072.00, 290.18) (140.79, 290.18) (140.79, 302.08) (072.00, 302.08)       /TT2 for each digest.  	<|special_separator|>
(072.00, 264.51) (359.32, 264.51) (359.32, 276.40) (072.00, 276.40)       /TT2 2. Use a micropipette to load the contents of each reaction tube (20 	<|special_separator|>
(359.41, 264.51) (364.86, 264.51) (364.86, 276.40) (359.41, 276.40)      /C2_1 𝜇𝜇	<|special_separator|>
(364.92, 264.51) (370.27, 264.51) (370.27, 276.40) (364.92, 276.40)       /TT4 L	<|special_separator|>
(370.20, 264.51) (527.06, 264.51) (527.06, 276.40) (370.20, 276.40)       /TT2  total) into a separate well in the gel. 	<|special_separator|>
(072.00, 252.74) (510.03, 252.74) (510.03, 264.64) (072.00, 264.64)       /TT2 Use a fresh pipet tip for each reaction tube and write down the order in which the samples are loaded.  	<|special_separator|>
(072.00, 227.07) (074.19, 227.07) (074.19, 238.96) (072.00, 238.96)       /TT2  	<|special_separator|>
(072.00, 201.27) (449.91, 201.27) (449.91, 213.16) (072.00, 213.16)       /TT2 NOTE: Be careful not to punch the tip of the pipet through the bottom or side of the well. 	<|special_separator|>
(072.00, 175.59) (074.19, 175.59) (074.19, 187.49) (072.00, 187.49)       /TT2  	<|special_separator|>
(072.00, 149.92) (137.55, 149.92) (137.55, 161.81) (072.00, 161.81)       /TT2 While loading,  	<|special_separator|>
(125.99, 127.24) (130.57, 127.24) (130.57, 135.74) (125.99, 135.74)      /C2_0 •	<|special_separator|>
(130.56, 126.53) (133.32, 126.53) (133.32, 135.90) (130.56, 135.90)       /TT1  	<|special_separator|>
(143.99, 123.51) (540.72, 123.51) (540.72, 135.41) (143.99, 135.41)       /TT2 steady the pipet over the well using two hands. You may wish to place one or both elbows on 	<|special_separator|>
(143.99, 111.87) (298.94, 111.87) (298.94, 123.76) (143.99, 123.76)       /TT2 the lab bench to steady your hands.  	<|special_separator|>
(125.98, 103.23) (130.56, 103.23) (130.56, 111.73) (125.98, 111.73)      /C2_0 •	<|special_separator|>
(130.54, 102.53) (133.31, 102.53) (133.31, 111.89) (130.54, 111.89)       /TT1  	<|special_separator|>
(143.98, 099.51) (536.76, 099.51) (536.76, 111.40) (143.98, 111.40)       /TT2 be careful to expel any air in the pipet tip end before loading the gel. If an air bubble forms a 	<|special_separator|>
(143.98, 087.86) (499.32, 087.86) (499.32, 099.76) (143.98, 099.76)       /TT2 cap over the well, the sample will flow into the buffer around the edges of the well.  