Team:Aachen/TestBench2

From 2014.igem.org

(Difference between revisions)
m (→LB medium)
m (→Hartmans minimal medium (HM))
Line 27: Line 27:
== Hartmans minimal medium (HM) ==
== Hartmans minimal medium (HM) ==
 +
 +
{| class="wikitable"
 +
|-
 +
! Component !! supplement for 1x HM [g/L] !! final 1x concentration
 +
|-
 +
| Casamino acids || 2 || 2 g/L
 +
|-
 +
| Thiamine hydrochloride || 0.3 || 300 mg/L
 +
|-
 +
| MgSO{{sub|4}}/MgSO{{sub|4}}*7H{{sub|2}}O || 0.242 / 0,494 || 2 mmol/L
 +
|-
 +
| CaCl{{sub|2}} || 0.011 || 0.1 mmol/L
 +
|}
= Agar Chips =
= Agar Chips =

Revision as of 22:47, 17 September 2014

Contents


Media

LB medium

  1. weight components
  • 5 g/L NaCl
  • 10 g/L tryptone
  • 5 g/L yeast extract
  • (15 g/L agar for plates)
  1. fill up to 1 L with deionized water
  • mix well by shaking
  1. autoclave
    1. autoclaving tape, caps slightly unscrewed
    2. base of the pot has to be covered with deionized water
    3. close lid
    4. heat level 3 until the pressure valve opens
    5. reduce heat level to 1.5
    6. set timer to 20 minutes
    7. turn heater off
    8. wait until the pressure valve retracts (30-45 minutes)
    9. open, close caps & shake
  2. for plates, wait until you can touch the bottle (<60 °C, clean bench!)
  3. add antibiotics and shake (gloves!)

Hartmans minimal medium (HM)

Component supplement for 1x HM [g/L] final 1x concentration
Casamino acids 2 2 g/L
Thiamine hydrochloride 0.3 300 mg/L
MgSO4/MgSO4*7H2O 0.242 / 0,494 2 mmol/L
CaCl2 0.011 0.1 mmol/L

Agar Chips

Transformation

Heat Shock

  1. thaw cells on ice
  2. add 1 µl of plasmid DNA
  3. incubate on ice for 30'
  4. heat shock at 42 °C for 60
  5. incubate on ice for 5'
  6. add 200 µl of SOC media
  7. incubate at 37 °C for 2 h
  8. plate 20  and 200 µl on plates with the appropiate antibiotic

Electroporation

PCR

Colony PCR

QuikChange

Clonings

Restriction Digest

Ligation

Gibson Assembly

Contact Disclaimer

Retrieved from "http://2014.igem.org/Team:Aachen/TestBench2"