Team:Aachen/Notebook/Wetlab/April
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AZimmermann (Talk | contribs) (Created page with "__NOTOC__ {{Team:Aachen/Header}} = April = == 1st == '''Organization''' where to get: * key cards and keys for the lab * cooled centifuge * space in -80 °C * bottles for ...") |
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** cells had been incubated in SOC or LB medium | ** cells had been incubated in SOC or LB medium | ||
+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
! Dilution !! 200 µl (stock) !! 100 µl (stock) !! 1:5 !! 1:10 !! 1:100 | ! Dilution !! 200 µl (stock) !! 100 µl (stock) !! 1:5 !! 1:10 !! 1:100 | ||
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|- | |- | ||
|} | |} | ||
+ | </center> | ||
* BL21 cells were centrifuged and plated; only 128 colonies grew in total indicating a very bad efficiency (~482.11). | * BL21 cells were centrifuged and plated; only 128 colonies grew in total indicating a very bad efficiency (~482.11). | ||
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'''Pcq lab PCR''' advanced primus 25, 96 | '''Pcq lab PCR''' advanced primus 25, 96 | ||
+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
! !! Lenght [bp] !! % GC !! T<sub>m</sub> !! T<sub>A</sub> | ! !! Lenght [bp] !! % GC !! T<sub>m</sub> !! T<sub>A</sub> | ||
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|- | |- | ||
|} | |} | ||
+ | </center> | ||
Program name IGEM.cyc | Program name IGEM.cyc | ||
+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
! Parameter !! Duration !! Temp [°C] | ! Parameter !! Duration !! Temp [°C] | ||
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| Store||forever||8 | | Store||forever||8 | ||
|} | |} | ||
+ | </center> | ||
# RFC25_F RFC25_R | # RFC25_F RFC25_R | ||
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* ca. 30 ng/µL | * ca. 30 ng/µL | ||
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{| class="wikitable" | {| class="wikitable" | ||
! !! 20x !! -> 20 µL thereof: 30x | ! !! 20x !! -> 20 µL thereof: 30x | ||
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| '''water'''||33.5||21.5 | | '''water'''||33.5||21.5 | ||
|} | |} | ||
+ | </center> | ||
'''Anneling''': 52 °C | '''Anneling''': 52 °C | ||
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*'''PCR with Phusion polymerase''' | *'''PCR with Phusion polymerase''' | ||
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{| class="wikitable" | {| class="wikitable" | ||
! !!volume [µL] | ! !!volume [µL] | ||
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| '''total'''||50 | | '''total'''||50 | ||
|} | |} | ||
+ | </center> | ||
*'''Digestion for restriction with NEB enzymes''' | *'''Digestion for restriction with NEB enzymes''' | ||
+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
! !!volume | ! !!volume | ||
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| '''total'''||??? µL | | '''total'''||??? µL | ||
|} | |} | ||
+ | </center> | ||
+ | |||
* ligation with NEB Kit | * ligation with NEB Kit | ||
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template A+B → 20 µL → <u>13.8 + 6.2</u> <u>5.3 + 14.7</u> | template A+B → 20 µL → <u>13.8 + 6.2</u> <u>5.3 + 14.7</u> | ||
+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
! !!volume [µL] | ! !!volume [µL] | ||
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| '''water'''||13.5 | | '''water'''||13.5 | ||
|} | |} | ||
+ | </center> | ||
* '''SOE2''' | * '''SOE2''' | ||
+ | <center> | ||
+ | [https://2014.igem.org/Team:Aachen/Notebook/Wetlab2 back to Wetlab main page] | ||
+ | </center> | ||
{{Team:Aachen/Footer}} | {{Team:Aachen/Footer}} |
Revision as of 15:04, 8 October 2014
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