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Edinburgh/20 June 2008
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* '''Result of transformation:''' the positive control was highly successful, with hundreds of colonies, but no colonies were present on the plate transformed with the BioBrick DNA indicating some problem with the DNA elution. Need to check that we got the method right. | * '''Result of transformation:''' the positive control was highly successful, with hundreds of colonies, but no colonies were present on the plate transformed with the BioBrick DNA indicating some problem with the DNA elution. Need to check that we got the method right. | ||
| - | * Ordered primers for ''dxs'', ''appY'' and ''glgC''. There are no forbidden restriction sites in the first two, so primers with full prefix and partial suffix were ordered to clone as EcoRI-SpeI fragments, but ''glgC'' has two EcoRI sites which will have to be mutated out. We therefore had a choice of cloning it initially as a XbaI-PstI fragment, or attempting the BABEL system, and decided to try BABEL first. If it proves too unreliable, we will need to order new primers to clone it the old-fashioned way. | + | * Ordered primers for ''dxs'', ''appY'' and ''glgC''. There are no forbidden restriction sites in the first two, so primers with full prefix and partial suffix were ordered to clone as EcoRI-SpeI fragments, but ''glgC'' has two EcoRI sites which will have to be mutated out. We therefore had a choice of cloning it initially as a XbaI-PstI fragment, or attempting the BABEL system, and decided to try BABEL first. If it proves too unreliable, we will need to order new primers to clone it the old-fashioned way. |
| - | '''Primer dxsf1:''' gat '''gaattc''' gcggccgc t '''tctaga''' tg agt ttt gat att gcc | + | ** '''Primer dxsf1:''' gat '''gaattc''' gcggccgc t '''tctaga''' tg agt ttt gat att gcc |
| - | '''Primer dxsr1:''' gc t '''actagt''' a tt a tta tgc cag cca ggc ctt g | + | ** '''Primer dxsr1:''' gc t '''actagt''' a tt a tta tgc cag cca ggc ctt g |
| - | '''Primer appyf1:''' gat '''gaattc''' gcggccgc t '''tctaga''' tg gat tat gtt tgc tcc | + | ** '''Primer appyf1:''' gat '''gaattc''' gcggccgc t '''tctaga''' tg gat tat gtt tgc tcc |
| - | '''Primer appyr1:''' gct '''actagt''' a tta tt a gtc aat tgt ttt gtt tat tcc | + | ** '''Primer appyr1:''' gct '''actagt''' a tta tt a gtc aat tgt ttt gtt tat tcc |
| - | '''Primer glgcf1:''' atg gtt agt tta gag aag aac gat c | + | ** '''Primer glgcf1:''' atg gtt agt tta gag aag aac gat c |
| - | '''Primer glgcr1:''' tta tta tcg ctc ctg ttt atg ccc taa c | + | ** '''Primer glgcr1:''' tta tta tcg ctc ctg ttt atg ccc taa c |
Revision as of 14:00, 20 August 2008
Week 1
Friday 20 June 08
- Result of transformation: the positive control was highly successful, with hundreds of colonies, but no colonies were present on the plate transformed with the BioBrick DNA indicating some problem with the DNA elution. Need to check that we got the method right.
- Ordered primers for dxs, appY and glgC. There are no forbidden restriction sites in the first two, so primers with full prefix and partial suffix were ordered to clone as EcoRI-SpeI fragments, but glgC has two EcoRI sites which will have to be mutated out. We therefore had a choice of cloning it initially as a XbaI-PstI fragment, or attempting the BABEL system, and decided to try BABEL first. If it proves too unreliable, we will need to order new primers to clone it the old-fashioned way.
- Primer dxsf1: gat gaattc gcggccgc t tctaga tg agt ttt gat att gcc
- Primer dxsr1: gc t actagt a tt a tta tgc cag cca ggc ctt g
- Primer appyf1: gat gaattc gcggccgc t tctaga tg gat tat gtt tgc tcc
- Primer appyr1: gct actagt a tta tt a gtc aat tgt ttt gtt tat tcc
- Primer glgcf1: atg gtt agt tta gag aag aac gat c
- Primer glgcr1: tta tta tcg ctc ctg ttt atg ccc taa c
