Team:Bordeaux/NotebookJune27

From 2014.igem.org

27th June

Molecular biology

  • IPTG
    Induction with 8ml IPTG
    m=0,19g qsp 8ml H2O for 1L

  • Oligos SLP
    We try to synthesize SLP gene with oligos

    Test with nanodrop :
    [SLP1]=1036,6ng/µl
    [SLP2]=1311,6ng/µl
    [SLP3]=1430,3ng/µl
    [SLP4]=1421,2ng/µl
    [SLP5]=1494,5ng/µl
    [SLP6]=1279,7ng/µl
    [SLP7]=1515,8ng/µl
    [SLP8]=541,4ng/µl

  • Protocol :
    - Oligo (10ng)
    - dNTP(1µl)
    - mix enzyme (15µl)

    We want a SLP concentration of 10ng/µl
    We do a mix and put it at 50°C


Production

  • ELP-20, -40 and -60 in LB+glucose
    Preculture inoculate the previous day


Purification

  • SDS-PAGE
    10µl P2 + 10µl sample 2X -> 5min at 85°C
    6µl ladder