Team:Stockholm/30 June 2010

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(Difference between revisions)
m (→Expression of SOD and yCCS from pEX expression vector)
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We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. '''Glycerol stocks''' were prepared from ON culture and '''pEX.SOD''' and '''pEX.yCCS plasmids''' were prepared.
We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. '''Glycerol stocks''' were prepared from ON culture and '''pEX.SOD''' and '''pEX.yCCS plasmids''' were prepared.
 +
 +
==== DNA concentrations ====
 +
{|
 +
|Sample
 +
|[nucleic acid] (ng/ul)
 +
|A(260)/A(280)
 +
|-
 +
|pEX.SOD
 +
|18.15
 +
|1.98
 +
|-
 +
|pEX.yCCS
 +
|20.55
 +
|1.97
 +
|}
 +
 +
*pEX.SOD:
== Preparation of competent BL21(DE3) cells ==
== Preparation of competent BL21(DE3) cells ==
* ON culture was set and grown in 37°C ON with 250 rpm rotary shaking.
* ON culture was set and grown in 37°C ON with 250 rpm rotary shaking.

Revision as of 14:00, 2 July 2010

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Contents

Morning meeting

We discussed the project proceedings.

  • It was decided that already amplified genes (cloned in pEX vector) should be transferred to pSB1C3 vector.
    • IgG protease
    • Superoxidase dismutase (SOD)
      • yCCS
    • bFGF
  • Primers for not-yet amplified genes should be redesigned to include the complete Assembly standard 25 prefix and suffix.
    • CPP
      • Transportan 10
      • LMWP
      • TAT
    • MITF
    • Protein A Z-domain
    • Tyrosinase
    • Vitamin B9 genes
  • BL21(DE3) was chosen as our strain for IPTG-induced protein expression from pEX vector.

Andreas

Expression of SOD and yCCS from pEX expression vector

Continued from 29/6

We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. Glycerol stocks were prepared from ON culture and pEX.SOD and pEX.yCCS plasmids were prepared.

DNA concentrations

Sample [nucleic acid] (ng/ul) A(260)/A(280)
pEX.SOD 18.15 1.98
pEX.yCCS 20.55 1.97
  • pEX.SOD:

Preparation of competent BL21(DE3) cells

  • ON culture was set and grown in 37°C ON with 250 rpm rotary shaking.
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