Team:Todai-Tokyo/Protocols/Notebook Sample
From 2009.igem.org
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<BR> | <BR> | ||
- | '''Results''':<BR> | + | '''Results''':<BR><BR> |
1) Sequence read failed | 1) Sequence read failed | ||
<BR>2) Single-base mutation found in sequence that creates stop codon; re-do the ligation | <BR>2) Single-base mutation found in sequence that creates stop codon; re-do the ligation | ||
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<h2> Infusion + Transformation </h2> | <h2> Infusion + Transformation </h2> | ||
- | The following constructs were created using [[Team:Todai-Tokyo/Protocols/Infusion|Infusion]] and the listed fragments: | + | The following constructs were created using [[Team:Todai-Tokyo/Protocols/Infusion|Infusion]] and the listed fragments, then [[Team:Todai-Tokyo/Protocols/Transformation|Transformed]] into ''E. coli'' competent cells: |
* pLacI-RBS-yqiT-dterm | * pLacI-RBS-yqiT-dterm | ||
- | ** pLacI-RBS '''S/P''' ([[Team:Todai-Tokyo/Protocols/Restriction Enzyme Digest|Digested]] on 10/1) | + | ** pLacI-RBS '''S/P''' ([[Team:Todai-Tokyo/Protocols/Restriction Enzyme Digest|Digested]] on 10/1) as vector |
- | ** yqiT-dterm ([[Team:Todai-Tokyo/Protocols/Miniprep|Miniprepped]] on 10/1) | + | ** yqiT-dterm ([[Team:Todai-Tokyo/Protocols/Miniprep|Miniprepped]] on 10/1) as insert |
+ | |||
+ | using the following primers:<BR> | ||
+ | * '''Inf fwd''': agtgtgtcgatgcagcccatgacacacacacagagatag | ||
+ | * '''Inf rev''': ggggagagatatatagagagacacacacagagatatat | ||
Latest revision as of 10:14, 20 October 2009
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このページの使い方
MiniprepThe following were miniprepped:
PCRThe yqiT gene was PCR amplified from pLacI-yqiT-dterm (miniprepped on 9/7) using the following primers: yqiT fwd: agcccgtgtagtactgtagagtt using PCR program 1 and Ex-taq. Note: This PCR reaction attaches a Biobrick prefix and suffix to the gene. *補足: Program 1と書いてあるところは、いつも使うようなプログラムと違うものを使ってそれを記したい場合はプログラムの内容をリンク先に書き込みましょう。わからない場合は聞いてください。
SequencingThe following were sequenced using the labeled primers:
Results:
TransformationThe following were Transformed into E. coli competent cells:
Ligation + TransformationThe following Ligations were performed using the listed fragments and Transformed into E. coli competent cells:
Infusion + TransformationThe following constructs were created using Infusion and the listed fragments, then Transformed into E. coli competent cells:
using the following primers:
RE Digest + Gel PurificationThe following were Digested using the listed Restriction Enzymes and then Gel Purified:
Colony PCRColonies from the following plates were used for Colony PCR using the listed primers:
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