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Revision as of 14:03, 2 October 2010 (view source) AndreasConstantinou (Talk | contribs) (New page: {{Stockholm/Top2}} ==Andreas==) ← Older edit		Revision as of 15:26, 2 October 2010 (view source) AndreasConstantinou (Talk | contribs) Newer edit →
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-	{{Stockholm/Top2}}
	==Andreas==		==Andreas==
		+	===Transfer of nCPP⋅SOD⋅His.RBS.yCCS operon to pEX===
		+	''Continued from 30/8 transformations''
		+	====Colony PCR====
		+	#<u>BL21</u> pEX.nLMWP&sdot;SOD&sdot;His: A & B
		+	#<u>BL21</u> pEX.nTra10&sdot;SOD&sdot;His: A & B
		+	#pEX.nTAT&sdot;SOD&sdot;His.RBS.yCCS 2: A & B
		+	#pEX.nTAT&sdot;SOD&sdot;His.RBS.yCCS 3: A & B
		+	#pEX.nTra10&sdot;SOD&sdot;His.RBS.yCCS 1: A & B
		+	#pEX.nTra10&sdot;SOD&sdot;His.RBS.yCCS 2: A & B
		+	#pEX.nLMWP&sdot;SOD&sdot;His.RBS.yCCS 2: A & B
		+	#pEX.nLMWP&sdot;SOD&sdot;His.RBS.yCCS 3: A & B
		+
		+	Standard colony PCR settings.
		+	*Elongation time: 2:00
		+
		+	====Gel verification====
		+	0.8 % agarose, 100 V
		+
		+	'''Expected bands:'''
		+	#744 bp
		+	#765 bp
		+	#1523 bp
		+	#1523 bp
		+	#1553 bp
		+	#1553 bp
		+	#1532 bp
		+	#1532 bp
		+
		+	'''Results'''<br />
		+	Ran gel a little too long too see correct clones of constructs '''1''' and '''2'''. For the other it looks like many clones have double inserts, probably a result of too high insert:vector ratio during ligation.
		+
		+	Confirmed clones:
		+	#-
		+	#-
		+	#A
		+	#B
		+	#-
		+	#-
		+	#B
		+	#B
		+
		+	These will be saved for later.

---

Revision as of 15:26, 2 October 2010

Contents 1 Andreas 1.1 Transfer of nCPP⋅SOD⋅His.RBS.yCCS operon to pEX 1.1.1 Colony PCR 1.1.2 Gel verification

Andreas

Transfer of nCPP⋅SOD⋅His.RBS.yCCS operon to pEX

Continued from 30/8 transformations

Colony PCR

1. BL21 pEX.nLMWP⋅SOD⋅His: A & B
2. BL21 pEX.nTra10⋅SOD⋅His: A & B
3. pEX.nTAT⋅SOD⋅His.RBS.yCCS 2: A & B
4. pEX.nTAT⋅SOD⋅His.RBS.yCCS 3: A & B
5. pEX.nTra10⋅SOD⋅His.RBS.yCCS 1: A & B
6. pEX.nTra10⋅SOD⋅His.RBS.yCCS 2: A & B
7. pEX.nLMWP⋅SOD⋅His.RBS.yCCS 2: A & B
8. pEX.nLMWP⋅SOD⋅His.RBS.yCCS 3: A & B

Standard colony PCR settings.

• Elongation time: 2:00

Gel verification

0.8 % agarose, 100 V

Expected bands:

1. 744 bp
2. 765 bp
3. 1523 bp
4. 1523 bp
5. 1553 bp
6. 1553 bp
7. 1532 bp
8. 1532 bp

Results
Ran gel a little too long too see correct clones of constructs 1 and 2. For the other it looks like many clones have double inserts, probably a result of too high insert:vector ratio during ligation.

Confirmed clones:

1. -
2. -
3. A
4. B
5. -
6. -
7. B
8. B

These will be saved for later.