Copenhagen/27 July 2011

From 2011.igem.org

(Difference between revisions)
 
Line 2: Line 2:
-
Der blev kørt koloni PCR på 10 tilfældige kolonier fra B1.
 
-
koloni 1,9 og 10 viste tegn på det rette insert.
 
-
Overnats kulturer fra 1,9 og 10 blev oprenset, en prøve blev digested med Ecor1 og PST1 for at se om det rigtige insert var der.
 
-
Der var 3 kolonier fra en transformation med en ligering (1 time på bord) Casper lavede af A2. De blev overført til en ny plade og hedder A,B,C. B viste tegn på insert og er sat til overnats opformering.
+
 
-
Casper havde lavet 2 ligeringer sammen med den før nævnte 1 times ligering, disse ligerede over nat.
+
 
-
De 2 overnats ligeringer blev transformeret.
+
 
'''Labwork'''
'''Labwork'''
Line 14: Line 11:
*Ligate the the two sets of plasmids with the insert and make two controls to test if the digestion went well. One with only T4 buffer and one with T4 buffer and T4 ligase.
*Ligate the the two sets of plasmids with the insert and make two controls to test if the digestion went well. One with only T4 buffer and one with T4 buffer and T4 ligase.
-
*Run colony pcr on 10 colonies from B1.
+
*Colony PCR was done to 10 randomly selected colonies of B1 to screen for one with the right insert. Colony 1,9 and 10 showed vague signs of having this insert.
 +
Overnight cultures of 1,9 and 10, which were made before knowing if they were any good, were purified and a sample from each was double digested with Ecor1 and PST1, whith the intention of determining whether or not the wanted insert was to be found.  
 +
*Yesterday Casper did 3 ligations of A2. One was transformed after a 1 hour table ligation period. 3 Colonies grew from this transformation, these were transferred to another plate and dubbed A,B and C. From a colony PCR colony B showed signs of having the insert. An overnight colture was made of B.
 +
The two other ligations which were overnight ligations were transformed.
Back to [[Team:Copenhagen/Notebook|Notebook]]
Back to [[Team:Copenhagen/Notebook|Notebook]]

Latest revision as of 13:25, 28 July 2011

Wednesday




Labwork

  • Ligate the the two sets of plasmids with the insert and make two controls to test if the digestion went well. One with only T4 buffer and one with T4 buffer and T4 ligase.
  • Colony PCR was done to 10 randomly selected colonies of B1 to screen for one with the right insert. Colony 1,9 and 10 showed vague signs of having this insert.

Overnight cultures of 1,9 and 10, which were made before knowing if they were any good, were purified and a sample from each was double digested with Ecor1 and PST1, whith the intention of determining whether or not the wanted insert was to be found.

  • Yesterday Casper did 3 ligations of A2. One was transformed after a 1 hour table ligation period. 3 Colonies grew from this transformation, these were transferred to another plate and dubbed A,B and C. From a colony PCR colony B showed signs of having the insert. An overnight colture was made of B.

The two other ligations which were overnight ligations were transformed.

Back to Notebook

Retrieved from "http://2011.igem.org/Copenhagen/27_July_2011"