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  • Bruno Vellutini 18:43 on 2012/04/13 Permalink
    Tags: , miniprep, , , , , , , ,   

    Katrine/Andi Terebratalia segmentation genes, miniprep specs 

    Since the previous sequencing for these minipreps did not work I am trying to sequence them again. These are the reads from the minipreps, they look ok.

    Gene ID ng/µL
    Tt gli 1 BV105 569.21
    Tt gli 2 BV106 755.79
    Tt gli 3 BV107 858.49
    Tt net 1 BV108 1080.2
    Tt net 2 BV109 416.45
    Tt net 3 BV110 452.18
    Tt net 3 BV111 474.99
    Tt nk6 1 BV112 1898.64
    Tt nk6 2 BV113 1217.41
    Tt nk6 3 BV114 1002.05
    Tt ptc 1 BV115 577.31
    Tt ptc 2 BV116 588.24
    Tt ptc 2 BV117 585.43
    Tt ptc 2 BV118 578.18
    Tt ptc 2 BV119 492.78
    Tt ptc 3 BV120 929.99
    Tt smo 1 BV121 631.31
    Tt smo 2 BV122 332.49
    Tt smo 3 BV123 1290.73
    Tt wnt7 1 BV124 968.39
    Tt wnt7 2 BV105 557.44
    Tt wnt7 3 BV106 2036.43
    Tt wnt7 4 BV107 554.82
    Tt wnt7 5 BV108 446.68

     
  • Bruno Vellutini 16:35 on 2012/01/15 Permalink
    Tags: , , miniprep, ,   

    MiniPrep for Membranipora MAEL F2, PUM, TUDOR 

    Picked positive colonies to grow. 9 samples on shake to grow at 16:30.

    16/01/12

    Sequencing reaction at 12:00 and tubes dropped by the sequencing facility at 15:00.

    ID Gene Vector Quality Probe Kit
    BV7 MAEL F2 T7  bad  –
    BV8 MAEL F2 T7  ok  SP6
    BV9 MAEL F2 T7  ok  SP6
    BV10 PUM T7  ok  SP6
    BV11 PUM T7  ok  T7
    BV12 PUM T7  ok  T7
    BV13 TUDOR T7  ok  T7
    BV14 TUDOR T7  ok  T7
    BV15 TUDOR T7  ok  T7
    BV16 MAEL F2 SP6  bad  –
    BV17 MAEL F2 SP6  ok  SP6
    BV18 MAEL F2 SP6  ok  SP6
    BV19 PUM SP6  ok  SP6
    BV20 PUM SP6  ok  T7
    BV21 PUM SP6  ok  T7
    BV22 TUDOR SP6  ok  T7
    BV23 TUDOR SP6  ok  T7
    BV24 TUDOR SP6  ok  T7

    19/01/12

    Sequences came back.

     
  • Bruno Vellutini 16:49 on 2012/01/09 Permalink
    Tags: , miniprep, ,   

    MiniPrep for Membranipora PIWIL2 and NANOS 

    09/01/12

    Two positive colonies form Mm PIWIL2 (1 and 3) and Mm NANOS (2 and 3) were picked and added to a growth tube with 3 mL of LB medium. Tubes were left in the S8 large shaker overnight at 210 rpm / 37 °C.

    10/01/12

    1.5 mL were poured to eppendorfs, centrifuged, discarded supernatant and followed the MiniPrep protocol. DNA was kept in the fridge overnight before the sequencing reaction.

    11/01/12

    PCR sequencing reaction and submitted samples:

    ID Gene Primer Quality Probe Kit
    BV1 PIWIL2 T7 bad
    BV2 PIWIL2 T7 bad
    BV3 NANOS T7 ok SP6
    BV4 NANOS T7 ok SP6
    BV5 PIWIL2 SP6 bad
    BV6 PIWIL2 SP6 short, ok SP6

    13/01/12

    Sequences came back, see above.

     
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