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Revision as of 10:00, 14 July 2011 (view source) Sandra.Wassner (Talk | contribs) (→Sequencing) ← Older edit		Revision as of 10:00, 14 July 2011 (view source) Sandra.Wassner (Talk | contribs) (→Sequencing) Newer edit →
Line 28:		Line 28:
	*H2O		*H2O
	*Seq. Buffer		*Seq. Buffer
-	*100ng DNA (Lys 1-Lys5)	+	*DNA (Lys 1-Lys5)
-	*2ng Primer	+	*Primer
	*Polymerase		*Polymerase
	<br/>		<br/>

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Revision as of 10:00, 14 July 2011

23. Labday

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Digestion of PCR products

Investigators: Julia

Digest of PCR product Ccas.

Ccas:

• Enzyme 1: XbaI
• Enzyme 2: PstI

in vector pSB1C3:

• Enzyme 1: SpeI
• Enzyme 2: PstI

Comments: RFP is not digested with SpeI and PstI. It was digested again, that time with NEB4 and XbaI.

Sequencing

Investigators: Theo

PCR of Lys1, Lys2, Lys3, Lys4, Lys5.

Materials:

• H2O
• Seq. Buffer
• DNA (Lys 1-Lys5)
• Primer
• Polymerase

PCR programm:

1. 95°C 3 min
2. 95°C 15 sec
3. 50°C 15 sec
4. 60°C 2 min
5. 4°C stored til needed again

step 2 to 4 was repeated 40x times.