Team:Todai-Tokyo/Protocols/Transformation

From 2009.igem.org

(Difference between revisions)
(New page: {{:Team:Todai-Tokyo/Template}} == Transformation Protocol == '''From iGEM Plates''' <BR> # Add 15µl TE (Tris-EDTA, pH 8.0) to well containing part and pipette up and down to resuspend...)
(→Transformation Protocol)
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# Take 1µl of the above solution and mix with partly thawed competent cells on ice
# Take 1µl of the above solution and mix with partly thawed competent cells on ice
# Leave on ice for 30 min.
# Leave on ice for 30 min.
-
# Heat Shock for at 42C for 45 seconds
+
# Heat Shock for at 42ºC  for 45 seconds
# Return eppendorf containing cells to ice and leave for 5 min.
# Return eppendorf containing cells to ice and leave for 5 min.
# Add 500µl LB and culture at 37C for 30 min.
# Add 500µl LB and culture at 37C for 30 min.
# Spread on plate with appropriate antibiotic resistance
# Spread on plate with appropriate antibiotic resistance
-
# Culture plate at 37C overnight
+
# Culture plate at 37ºC  overnight
 +
<BR><BR>
 +
'''After ligation or from miniprep'''
-
'''After ligation or from miniprep'''<BR>
+
# Take 1µl (if from miniprep)/5µl (if from ligation) of DNA solution and mix with partly thawed competent cells on ice
-
 
+
-
 
+
-
# Take 1µl(if from miniprep)/5µl(if from ligation) of DNA solution and mix with partly thawed competent cells on ice
+
# Leave on ice for 30 min.
# Leave on ice for 30 min.
-
# Heat Shock for at 42C for 45 seconds
+
# Heat Shock for at 42ºC  for 45 seconds
# Return eppendorf containing cells to ice and leave for 5 min.
# Return eppendorf containing cells to ice and leave for 5 min.
-
# Add 500µl LB and culture at 37C for 30 min.
+
# Add 500µl LB and culture at 37ºC  for 30 min.
# Spread on plate with appropriate antibiotic resistance
# Spread on plate with appropriate antibiotic resistance
-
# Culture plate at 37C overnight
+
# Culture plate at 37ºC  overnight

Revision as of 06:46, 20 October 2009

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Transformation Protocol

From iGEM Plates

  1. Add 15µl TE (Tris-EDTA, pH 8.0) to well containing part and pipette up and down to resuspend
  2. Take 1µl of the above solution and mix with partly thawed competent cells on ice
  3. Leave on ice for 30 min.
  4. Heat Shock for at 42ºC for 45 seconds
  5. Return eppendorf containing cells to ice and leave for 5 min.
  6. Add 500µl LB and culture at 37C for 30 min.
  7. Spread on plate with appropriate antibiotic resistance
  8. Culture plate at 37ºC overnight



After ligation or from miniprep

  1. Take 1µl (if from miniprep)/5µl (if from ligation) of DNA solution and mix with partly thawed competent cells on ice
  2. Leave on ice for 30 min.
  3. Heat Shock for at 42ºC for 45 seconds
  4. Return eppendorf containing cells to ice and leave for 5 min.
  5. Add 500µl LB and culture at 37ºC for 30 min.
  6. Spread on plate with appropriate antibiotic resistance
  7. Culture plate at 37ºC overnight
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