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Team:Queens-Canada/12 August 2010
From 2010.igem.org
(New page: <h2>August 12, 2010</h2> {{:Team:Queens-Canada/head}} <h3>PCR</h3> Hao and Thai '''pGpd-2''', '''pHsp-3''' Program run using 9:1 FAST:HIFI blend wit...) |
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'''ChR2'''::'''eYFP'''::'''unc-54 3'UTR'''::1T3, '''ChR2'''::'''eYFP'''::'''unc-54 3'UTR'''::1T3, '''ChR2'''::'''eYFP'''::1C3, '''eGFP'''::1C3 | '''ChR2'''::'''eYFP'''::'''unc-54 3'UTR'''::1T3, '''ChR2'''::'''eYFP'''::'''unc-54 3'UTR'''::1T3, '''ChR2'''::'''eYFP'''::1C3, '''eGFP'''::1C3 | ||
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| + | <h3>[[Team:Queens-Canada/transformation|Microinjection]]</h3> | ||
| + | Yuli | ||
| + | |||
| + | Prepared injection pads. | ||
{{:Team:Queens-Canada/foot}} | {{:Team:Queens-Canada/foot}} | ||
Latest revision as of 02:02, 28 October 2010
August 12, 2010
PCR
Hao and Thai
pGpd-2, pHsp-3
Program run using 9:1 FAST:HIFI blend with an extension time of 25 s and annealing temperature of 57 °C.
Heat Shock
Hao and Thai Digestion-Ligation products from 11/08/10, unc-54 3'UTR::1A3
Used Amp plates for unc-54 3'UTR::1A3 and Chlor plates for all others.
Digestion
Chris
pStr-220::1C3
Cut with EcoRI and PstI.
Digestion-Ligation
Yuli and Steve
ChR2, unc-54 3'UTR, pSB1T3, pSB1C3, ChR2::YFP, eGFP
unc-54 3'UTR cut with XbaI and PstI. ChR2::YFP cut with EcoRI and SpeI. All others cut with EcoRI and PstI.
Digestion products ligated into 1T3 and 1C3.
Gel Electrophoresis
Hao
pStr-220, pHsp-3, pGpd-2, ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::1C3, 1T3, 1C3, eGFP::1C3
Used 0.7 g agarose in 100 mL TBE. Gel run for 60 mins at 100 V.
Heat Shock
Yuli and Steve
ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::1C3, eGFP::1C3
Microinjection
Yuli
Prepared injection pads.
