Team:Shenzhen SZMS/Lab

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Date Event Results Problem summary
Apr.3 B0015(chl) B0030(chl) Plasmid Transformation Done (yet this B0030 is single error)
Apr.4 Single clone selected from the work on Apr.3 (In order to be pithy, this process is omitted from the table in all the following daily work) Done
Apr.5 Plasmid preped and culture restored from the bacterial suspension on Apr.4 (omitted later for pithiness and convenience as well) Done
Apr.6 K115016(CHL) K115017(CHL) C0051(CHL) transformation Done
Apr.2 J23106(AMP) K082003(CHL,as GFP+LVA) transformation (help offered by Mentor Wang) Done
May.5 Digestion: K115016(S/P) C0051(X/P) C0051(E/S) B0015(E/X)
Electrophoresis
Gel extraction
16sheshidu℃ overnight
Done
May.6 Product from 5.5(CHL)transformed Failed Might be the problem during the plasmid prep
May.7 Plasmid mentioned above repreped Done
May.13 Digestion: C0051(E/S) B0015(S/P) K082003(E/S)
Electrophoresis
Gel extraction
16℃ overnight
Done, C0051-B0015
K082003-B0015 gained
May.14 Products from 5.13(CHL)all transformed Done, C0051-B0015
K082003-B0015 gained
May.15 Gel confirmation (products from 5.14), comparison with gel of B0015 Done, product longer than the original plasmid
May.27 Digestion: C0051-B0015(X/P) K115016(S/P) K115017(S/P)
Electrophoresis
Gel extraction
Combination
Transformation(CHL)
Done,
K115016-C0051-B0015
K115017-C0051-B0015
gained.
May.28 B0030(AMP) transformation Done; informed that R0051 requires PCR in order to get the primer of EC 1.7.7.2
May.29 Digestion: J23106(S/P)
K115016-C0051-B0015(X/P)
K115017-C0051-B0015(X/P)
Electrophoresis
Gel extraction
Combination
Transformation(AMP)
Done,
J23106-K115016-C0051-B0015(27℃CI-protein)
J23106-K115017-C0051-B0015(32℃CI-protein)
gained.
May.30 The result of sequencing of C0051-B0015
K082003-B0015 received
Correctly combined
Jun.2 Digestion: B0030(S/P) K082003-B0015(X/P)
Electrophoresis
Gel extraction
Combination
Transformation(AMP)
Done
Jun.8 The results of sequencing of J23106-K115016-C0051-B0015(27℃CI) and J23106-K115017-C0051-B0015(32℃ CI)received. Correctly combined
Jun.9 Digestion:B0030-K082003-B0015(E/S)
J23106-K115017-C0051-B0015(E/X)
Electrophoresis
Gel extraction
Combination
Transformation(AMP)
Done with transformation;
Yet failed in the electrophoretic analysis.
Disconnected
Jun.10 ciF1 ciF2 ciF3: primers to synthesize EC 1.7.7.2 and PEP (additional PEP R0051-B0030-) gained.
Jun.11 With the original blue 2X Master Mix PCR round 1,2,3 implemented Failed; only primer founded in the electrophoretic analysis
Jun.12 Round 1 from 6.11 repeated in the morning;
Round 3 PCR with 2X hi-fi Master Mix at noon
Failed in the morning but done in the afternoon;
R0051-B0030-1.7.7.2 recycled product gained.
might be the problem of MIX
Jun.13 Digestion: 27℃ CI(S/P)R0051-B0030-1.7.7.2(X/P)
32℃ CI(E/X)
B0030-K082003-B0015(E/S)
Electrophoresis
Gel
extraction
Combination
Transformation