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console.js:26

Tuesday - July 19

Monday, 7/18

Members Present: Tam, Kat, Marc, Karim, Hamed

LAB:

Morning:

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Miniprepped yesterday's cultures (16 in total)

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Conducted PCR on Backbone:

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Using RFP plasmid as template (x4)

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Using iGEM linearized backbone from the kit (x3)

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Conducted gradient PCR on Long GolS P118A construct

Afternoon:

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Made 3x200mL BL21 cultures in TB (left in rm315 shaker)

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Nanodropped the miniprepped cultures:

	A	B	C	D	E
1		Sample	260/280	260/230	ng/uL
2	1	AmilCP	1.9	0.99	15.4
3	2	L2_B	1.73	0.81	11.2
4	3	L2_A2	1.61	0.77	11.7
5	4	L3_A	1.89	0.59	10.5
6	5	L4_A1	1.59	0.99	14.3
7		DH10B + Amil	2.39	0.51	5.2
8	6	L5_B	1.8	0.96	16.5
9	7	Cj Blue	1.9	1.01	17.3
10	8	L2_A4	1.87	0.96	13.1
11	9	L5_A	1.71	0.74	10.5
12	10	L2 Red (Labelled 22 Red)	1.77	0.93	18
13		Cj Blue DH10B	1.59	0.46	7.3
14	11	L4_A2	1.71	1	11.6
15	12	L2_A2	1.78	0.97	18.3
16	13	L2_A3	1.66	0.76	9.9
17	14	L2_Red	1.76	0.93	17.4

Table1

L2: mCherry; L3: Long Linear P118A; 4: Long Linear GolS; 5: TetO_LacZ (all selected colonies were white aside from those marked red; AmilCP and Cj Blue were directly put into liquid LB+CAM post transformation)

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Digested miniprepped products using PstI (20ul reaction for running the gel only)

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Ran a gel on:

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All 7 backbone PCR products:

Gel1.jpg

Lane 1-3: PCR product of backbone using RFP plasmid as template Lane 4: Ladder Lane 5: PCR product of backbone using RFP plasmid as template Lane 6-8 PCR Product of backbone using iGEM linearized backbone from kit

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Gradient PCR of Long GolS P118A:

Gel2.jpg

Lanes 1-3: Gradient PCR products from highest temperature to the lowest (72, 71.4, 70) Lane 4: Ladder Lanes 5-6: Gradient PCR products from highest temperature to the lowest (67.6, 64.7) [Please see: https://www.neb.com/faqs/1/01/01/why-are-there-low-molecular-weight-discrete-bands-on-an-agarose-gel-after-a-pcr-using-phusion-reg]

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Digested miniprepped products:

Gel1_(miniprep).jpg

1 AmilCP 2 L2_B (white) 3 Ladder 4 L2_A2 (white) 5 L3_A 6 L4_A1 7 L5_B 8 Cj Blue

Gel2_(miniprep).jpg

1 L2_A4 (white) 2 L5_A 3 Ladder 4 L2 Red (Labelled 22 Red) 5 L4_A2 6 L2_A2 (white) 7 L2_A3 (white) 8 L2_Red

Administrative:

	A	B	C	D	E	F
1	Name	SKU	Unit Price	Quantity	Total (incl. tax)	Status
2	Shrimp Alkaline Phosphatase (rSAP) - 500 units	M0371S	71.68	1	81	available
3	Allience Gloves S	211-6205	7.5	2	16.95	available
4	Allience Gloves M	211-6206	7.5	2	16.95	available
5	Allience Gloves L	211-6207	7.5	2	16.95	available
6	Allience Gloves XL	211-6208	7.5	2	16.95	available

Table2