Minnesota/27 July 2008

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Revision as of 22:26, 27 July 2008

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1. Spec the plasmid prep products: Place 10 samples into 10 separate plate wells. 11 wells total were used: 1st well was the control so it had 36uL of distilled water and 4uL of elution buffer (since want everything except DNA to be control group), then 2-11 wells had 1-10 plasmid prep products. 2-11 wells had 36uL of distilled water added and 4uL of plasmid prep products. Results:

a. (1) Lac I #1 = [DNA] ng/uL is 45, [DNA] ug/uL is 0.045 HIGH/GOOD

b. (2) Lac I = [DNA] ng/uL is 35, [DNA] ug/uL is 0.035 HIGH/GOOD

c. (3) Lac I = [DNA] ng/uL is 35, [DNA] ug/uL is 0.035 HIGH/GOOD

d. (4) BV:Lac #2 = [DNA] ng/uL is 10, [DNA] ug/uL is 0.001 LOW/POOR

e. (5) BV:Tet #1 = [DNA] ng/uL is 0, [DNA] ug/uL is 0.00 NONE/BAD

f. (6) BV:Tet #2 = [DNA] ng/uL is 5, [DNA] ug/uL is 0.005 LOW/POOR

g. (7) BV:Tet/p22 #1 = [DNA] ng/uL is 5, [DNA] ug/uL is 0.005 LOW/POOR

h. (8) BV:Tet/p22 #1 = [DNA] ng/uL is 5, [DNA] ug/uL is 0.005 LOW/POOR

i. (9) BV:Tet/p22 #2 = [DNA] ng/uL is 335, [DNA] ug/uL is 0.335 HIGH/GOOD

j. (10) BV:Tet/p22 #2 = [DNA] ng/uL is 15, [DNA] ug/uL is 0.015 LOW/POOR

2. Digest Rxn: Using #'s: 1, 2, 6, 9, and 10 from above. Also using: p22cII, LAMBDAcI, and RFP from different plasmid prep days. After following table below, allow to incubate for 2 hours. Heat inactivate enzymes @ 65C for 15 minutes in water bath. Follow the table below:

Parts	10x Buffer	BSA	DNA	RE 1	RE 2	H20
BV:Lac I (1)	5.0uL	0.5uL	22.0uL	Spe1, 1.0uL	Pst1, 1.0uL	20.5uL
BV:Lac I #2 (2)	5.0uL	0.5uL	28.5uL	Spe1, 1.0uL	Pst1, 1.0uL	14.0uL
BV:Tet (6)	5.0uL	0.5uL	35.0uL	Spe1, 1.0uL	Pst1, 1.0uL	7.5uL
BV:Tet/p22 #1 (9)	5.0uL	0.5uL	3.0uL	Spe1, 1.0uL	Pst1, 1.0uL	39.5uL
BV:Tet/p22 #2 (10)	5.0uL	0.5uL	33.0uL	Spe1, 1.0uL	Pst1, 1.0uL	9.5uL
p22cII	5.0uL	0.5uL	5.0uL	Xba1, 1.0uL	Pst1, 1.0uL	37.5uL
LAMBDAcI	5.0uL	0.5uL	7.0uL	Xba1, 1.0uL	Pst1, 1.0uL	35.5uL
RFP	5.0uL	0.5uL	4.0uL	Xba1, 1.0uL	Pst1, 1.0uL	38.5uL

@@ Line 30: / Line 30: @@
 |'''j.''' (10) BV:Tet/p22 #2 = [DNA] ng/uL is 15, [DNA] ug/uL is 0.015    LOW/POOR
 |-
-|'''2. Digest Rxn:''' Using #'s: 1, 2, 6, 9, and 10 from above. Follow the table below:
+|'''2. Digest Rxn:''' Using #'s: 1, 2, 6, 9, and 10 from above. Also using: p22cII, LAMBDAcI, and RFP from different plasmid prep days. After following table below, allow to incubate for 2 hours. Heat inactivate enzymes @ 65C for 15 minutes in water bath. Follow the table below: