Team:Stockholm/22 September 2010

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(→Andreas)
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::'''***Note to self***'''<br />
::'''***Note to self***'''<br />
::''It might be possible that pSB1A2.RBS&sdot;yCCS clones have been mixed up with pEX.N-TAT&sdot;SOD&sdot;His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.
::''It might be possible that pSB1A2.RBS&sdot;yCCS clones have been mixed up with pEX.N-TAT&sdot;SOD&sdot;His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.
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== Mimmi ==
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=== SOD / SOD.his / his.SOD / yCCS ===
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{|
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| colspan="3" | ''colony PCR''
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| rowspan="8" width="100" |
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! samples
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| rowspan="8" width="100" |
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! colspan="2" | Conditions
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| rowspan="3" |
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|-
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| colspan="3" |
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| pEX.SOD x3
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! time
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! &deg;C
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|-
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! mix
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| (µl)
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| x13
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| pEX.SOD.his x3
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| 5m
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| 95
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|-
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| mastermix
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| 24.5
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| rowspan="5" |
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| pEX.his.SOD x3
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| 30s
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| 95
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| )
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|-
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| DNA
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| 0.5
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| pEX.yCCS x3
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| 30s
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| 55
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| > 30 cycles
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|-
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| align="right" | tot
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| 25µl
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| rowspan="3" |
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| 1m20s
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| 72
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| )
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|-
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| colspan="2" rowspan="2" |
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| 5m
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| 72
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|-
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| oo
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| 25
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|}
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==== Gel ====
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[[Image:place_for_picture.jpg|200px|thumb|left|]]
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{|
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! well
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! width="120" | sample
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! well
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! sample
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|-
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| 1
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| ladder
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| 9
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| ladder
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|-
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| 2
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| pEX.SOD.his 1
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| 10
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| pEX.SOD 1
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|-
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| 3
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| pEX.SOD.his 2
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| 11
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| pEX.SOD 2
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|-
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| 4
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| pEX.SOD.his 3
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| 12
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| pEX.SOD 3
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|-
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| 5
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| pEX.his.SOD 1
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| 13
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| pEX.yCCS 1
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|-
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| 6
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| pEX.his.SOD 1
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| 14
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| pEX.yCCS 2
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|-
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| 7
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| pEX.his.SOD 1
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| 15
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| pEX.yCCS 2
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|-
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| 8
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| pEX.SOD.his
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| 16
 +
| -
 +
|}

Revision as of 13:01, 4 October 2010

Home Project Idea Lab Work Results Modelling Team       Follow us on and        2010.igem.org

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Contents

Andreas

Gel verification

Colony PCR gel verification of new assemblies: pSB1A2.RBS.yCCS, pEX.N-TAT*SOD*His, pSB1C3.N-LMWP*SOD*His & pSB1K3.N-LMWP*SOD*His.
4 μl λ; 4 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.

Of 21/9 colony PCR samples

1 % agarose, 120 V

Expected bands

  • pSB1A2.RBS⋅yCCS (pA.RBSy): 1018 bp
  • pEX.N-TAT⋅SOD⋅His (pEX.nTSH): 735 bp
  • pSB1C3.N-LMWP⋅SOD⋅His (pC.nLSH): 857 bp
  • pSB1K3.N-LMWP⋅SOD⋅His (pK.nLSH): 857 bp
  • pSB1C3.SOD⋅His (PC): 815 bp

Results

  • pSB1A2.RBS⋅yCCS: Seemingly correct-sized bands for clones 2, 3 and 4.
  • pEX.N-TAT⋅SOD⋅His: Seemingly correct-sized bands for clones 3 & 4.
  • pSB1C3.N-LMWP⋅SOD⋅His: Seemingly correct-sized bands for clones 1 & 4.
  • pSB1K3.N-LMWP⋅SOD⋅His: Seemingly correct-sized band for clone 1.

ON cultures

  1. 5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 °C, 225 rpm
    • pSB1A2.RBS⋅yCCS: clones 3 and 4
    • pEX.N-TAT⋅SOD⋅His: clones 3 and 4
    • pSB1C3.N-LMWP⋅SOD⋅His: clones 1 and 4
    • pSB1K3.N-LMWP⋅SOD⋅His: clone 1
***Note to self***
It might be possible that pSB1A2.RBS⋅yCCS clones have been mixed up with pEX.N-TAT⋅SOD⋅His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.




Mimmi

SOD / SOD.his / his.SOD / yCCS

colony PCR samples Conditions
pEX.SOD x3 time °C
mix (µl) x13 pEX.SOD.his x3 5m 95
mastermix 24.5 pEX.his.SOD x3 30s 95 )
DNA 0.5 pEX.yCCS x3 30s 55 > 30 cycles
tot 25µl 1m20s 72 )
5m 72
oo 25


Gel

Place for picture.jpg
well sample well sample
1 ladder 9 ladder
2 pEX.SOD.his 1 10 pEX.SOD 1
3 pEX.SOD.his 2 11 pEX.SOD 2
4 pEX.SOD.his 3 12 pEX.SOD 3
5 pEX.his.SOD 1 13 pEX.yCCS 1
6 pEX.his.SOD 1 14 pEX.yCCS 2
7 pEX.his.SOD 1 15 pEX.yCCS 2
8 pEX.SOD.his 16 -
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