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Revision as of 12:23, 26 June 2013 (view source) Helencook (Talk | contribs) (→208 lab) ← Older edit		Revision as of 07:24, 27 June 2013 (view source) KrDa (Talk | contribs) (→Main purposes today) Newer edit →
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	*25 rxns including duplicates		*25 rxns including duplicates
		+
		+	*34 rxna including duplicates with the following setup:
		+	Tube 1-6 containing 5uL pZA21 plasmid template and 3uL of each primers for amplification of the backbone. Tube 7-10; 1uL g-Block from previous purification and 3uL of each Sec signal primers. Tube 11-14 containing same amount of g-Block and primers but now replaced with the TAT2 signal primers. Tube 15-18; 5uL GFP SF plasmid template and 3uL of each primers for GFP SF TAT. Tube 19-22; 5uL GFP SF plasmid template and 3uL of each primers for GFP SF Sec. Tube 23-26; 5uL RFP plasmid template and 3uL of each primer for RFP. Tube 27-30; 1uL g-Block purification and 3uL of each TAT3 primers. Tube 31-34; 0.5uL of the original g-Block and 3uL of each primers for amplifying the g-Block.
		+	All tube with less than 5uL template DNA was filled with MQ until final volume of 11uL(5+3+3).
		+
		+	First half of the samples where run on PCR with x7-polymerase and the second half where run with PHUSION-polymerase, with the exception of the g-Block where all reactions where run with PHUSION. This gives following setup:
		+	x7-poly in tube:
		+	*1-3, 7-8, 11-12, 15-16, 19-20, 23-24, 27-28
		+	PHUSION in tube:
		+	*4-6, 9-10, 13-14, 17-18, 21-22, 25-26, 29-30, 31-34
		+
		+	A mastermix was made for both PCR with
	==Who was in the lab==		==Who was in the lab==

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Revision as of 07:24, 27 June 2013

Contents 1 208 lab 1.1 Main purposes today 1.2 Who was in the lab 1.3 Procedure 1.4 Conclusion from today

208 lab

Main purposes today

helloworld-project

• 25 rxns including duplicates

• 34 rxna including duplicates with the following setup:

Tube 1-6 containing 5uL pZA21 plasmid template and 3uL of each primers for amplification of the backbone. Tube 7-10; 1uL g-Block from previous purification and 3uL of each Sec signal primers. Tube 11-14 containing same amount of g-Block and primers but now replaced with the TAT2 signal primers. Tube 15-18; 5uL GFP SF plasmid template and 3uL of each primers for GFP SF TAT. Tube 19-22; 5uL GFP SF plasmid template and 3uL of each primers for GFP SF Sec. Tube 23-26; 5uL RFP plasmid template and 3uL of each primer for RFP. Tube 27-30; 1uL g-Block purification and 3uL of each TAT3 primers. Tube 31-34; 0.5uL of the original g-Block and 3uL of each primers for amplifying the g-Block. All tube with less than 5uL template DNA was filled with MQ until final volume of 11uL(5+3+3).

First half of the samples where run on PCR with x7-polymerase and the second half where run with PHUSION-polymerase, with the exception of the g-Block where all reactions where run with PHUSION. This gives following setup: x7-poly in tube:

• 1-3, 7-8, 11-12, 15-16, 19-20, 23-24, 27-28

PHUSION in tube:

• 4-6, 9-10, 13-14, 17-18, 21-22, 25-26, 29-30, 31-34

A mastermix was made for both PCR with

Who was in the lab

Henrike, Jakob, Kristian

Procedure

PCR for 7 fragments:

Conclusion from today