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min

(Science: abbreviation) minute.


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Annealing Temparature

... rest at 65°C. Denaturation for 10s will be enough, but a bit higher temperature 95-98°C might be better. Elongation depends on polymerase, so 1 min can be enough for Phusion, but 4 min will be required for Pfu.

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by vladCH
Sat Feb 22, 2014 7:54 pm
 
Forum: Molecular Biology
Topic: Annealing Temparature
Replies: 4
Views: 1461

Annealing Temparature

... to have big difference in the Tms, second the Tms seem to be quite high (62 - 69°C). Are you sure your denaturation in the beginning is only 1 min or is it a typo? Why do you have such long denaturation during cycling? Why do you have such long annealing? What polymerase do you use? What is ...

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by JackBean
Tue Feb 18, 2014 12:40 pm
 
Forum: Molecular Biology
Topic: Annealing Temparature
Replies: 4
Views: 1461

Re: centrifugation of hemocytes

I can't see any pellet, BUT today it tried again, after 24h of incubation (cells have time to adhere), and centrifugation at 400 for 5 min, I gently took off the medium and resuspend the cells by pipetting thoroughly up/down: the viability worked well. However, that time i skiped the washing ...

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by Thom
Tue Jan 28, 2014 4:16 pm
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 2601

centrifugation of hemocytes

... a 96-well plate. Theses cells are circulating blood cells but display quite good adherence. Here is my problem: after centrifugation ( I tried 5 min at 300g, 10 min at 300g and 5 min at 1000g, in eppendorf and with 96-well plates, flat-bottom and round bottom). I took the supernatant off and ...

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by Thom
Mon Jan 27, 2014 6:41 pm
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 2601

Problems with FluoroJade staining

... dry overnight. This happens at step 3 of the protocol, I will slash all the variations I've found on the internet and tried with no success: 1) 5 min 80% ethanol, 20% NaOH/ 5 min 100% ethanol/ 3 min 80% ethanol 2) 2 minutes 70% ethanol/ 1 min 70% Ethanol 3) 1 minute dH2O (here is where the wrinkles ...

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by fabulousparrot
Mon May 20, 2013 1:47 pm
 
Forum: Molecular Biology
Topic: Problems with FluoroJade staining
Replies: 0
Views: 1537
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