Team:TU Delft/8 June 2010 content

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Revision as of 09:07, 17 July 2010

Pieter with eppy

Eppy

Pieter just got a little present from Mariska van Ham :-) He's so happy with is eppy!

Today Nadine & Luke are learning how to work with HTML.

Lab work

Thias PCRed some BioBricks with various primer concentrations and at different temperatures to see which PCR method would work best. The PCR products were tested on a gel, and we saw a difference between the primer concentrations and the temperatures used.

BioBrick stocks

A number of Biobricks from the DNA distribution plates were suspended yesterday . Hugo and Kira did a transformation with 2 μL of the following BioBricks (antibiotic):

pSB3T5 (Tetracycline)
pSB1T3 (Tetracycline)
pSB1C3 (Chloramphenicol)
pSB3C5 (Chloramphenicol)
pSB1K3 (Kanamycin)
pSB1A3 (Ampicillin)
I13401 (Ampicillin)

Because our own competent cells were not good enough we used commercially available competent Top10 cells (Invitrogen). The cells were plated on LB-agar with the appropriate antibiotic. Biobricks plated We will have to wait until tomorrow to see if they were successful or not. Hopefully we will see some colonies tomorrow. Wish us luck!

@@ Line 12: / Line 12: @@
 Thias [[Team:TU_Delft/protocols/PCR|PCRed]] some BioBricks with various primer concentrations and at different temperatures to see which PCR method would work best. The PCR products were tested on a gel, and we saw a difference between the primer concentrations and the temperatures used.
-Hugo and Kira did a [[Team:TU_Delft/protocols/transformation|transformation]]:
+<h4>BioBrick stocks</h4>
-A number of Biobricks from the DNA distribution plates were resuspended and added to commercial competent Top10 cells. 15μl of the competent cells was added to 2μl of suspended DNA and incubated on ice for 30 min. This was followed by a heat-shock at 42°C (waterbath) for 30 seconds, addition of 50μl of sterile SOC, and an 1 hour incubation at 37°C. The cells were plated on LB-agar with the appropriate antibiotic.
+A number of Biobricks from the DNA distribution plates were suspended  [https://2010.igem.org/Team:TU_Delft#/blog?blog=7_June_2010  yesterday] . Hugo and Kira did a  [[Team:TU_Delft/protocols/transformation2| transformation]] with 2 μL of the following BioBricks (antibiotic):
-Biobricks plated (antibiotic):
+* pSB3T5 (Tetracycline)
-* pSB3T5 (T)
+* pSB1T3 (Tetracycline)
-* pSB1T3 (T)
+* pSB1C3 (Chloramphenicol)
-* pSB1C3 (C)
+* pSB3C5 (Chloramphenicol)
-* pSB3C5 (C)
+* pSB1K3 (Kanamycin)
-* pSB1K3 (K)
+* pSB1A3 (Ampicillin)
-* pSB1A3 (A)
+* I13401 (Ampicillin)
-* GFP-TT (A)
-* Top10 without biobrick (to test the cells) (-)
-We will have to wait until tomorrow to see if they were successful or not.
+Because our own competent cells were not good enough we used commercially available competent Top10 cells (Invitrogen). The cells were plated on LB-agar with the appropriate antibiotic. Biobricks plated
+We will have to wait until tomorrow to see if they were successful or not. Hopefully we will see some colonies tomorrow. Wish us luck!