Team:Hong Kong HKUST/notebook/mod3
From 2013.igem.org
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<div> | <div> | ||
<a href='#' class='head'>Week 1</a> | <a href='#' class='head'>Week 1</a> | ||
- | <div class='content' align="left"> | + | <div class='content' align="left"> |
+ | <p> </p> | ||
+ | <p><strong>August 5</strong></p> | ||
+ | <p>GFP (25)PCR Check with VR VF2</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Gel check(overan)</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>BBa_J176171</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Digestion using SpeI and PstI-HF to extract the backbone</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>PolyA / BBa_K404108</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Digestion using XbaI and PstI-HF</p> | ||
+ | <p>- Gel electrophoresis</p> | ||
+ | <p>- Run Gel </p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>Inoculation of pSB1C3</p> | ||
+ | <p> </p> | ||
+ | <p><strong>August 6</strong></p> | ||
+ | <p>GFP (25)</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Repeat PCR check</p> | ||
+ | <p>- Run Gel</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>pSB1C3</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Plasmid extraction</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>PolyA / BBa_K404108</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Gel purification</p> | ||
+ | <p>- Gel check</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>BBa_J176171</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- PCR clean up</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>BBa_K929000 & BBa_J52034 </p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Transformation using CAM plate</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>MLS</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- PCR amplify MLS in RFC 10 and 25 format</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p><strong>August 7</strong></p> | ||
+ | <p>MLS (RFC 10 & 25)</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- PCR clean up</p> | ||
+ | <p>- Nanodrop for the PCR clean up product</p> | ||
+ | <p>- Gel check for the PCR clean up product</p> | ||
+ | <p>- Digestion using EcoRI and PstI</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>PolyA / BBa_K404108</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Gel check</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>pSB1C3 </p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Digest with EcoRI & PstI </p> | ||
+ | <p>- Run gel and cut gel </p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>GFP(25) </p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Digestion with EcoRI & SpeI</p> | ||
+ | <p>- Run gel and cut gel </p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p><strong>August 8</strong></p> | ||
+ | <p>GFP25 & pSB1C3</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Gel purification</p> | ||
+ | <p>- Nanodrop</p> | ||
+ | <p>- Gel check </p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>polyA</p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Digestion with EcoRI and XbaI</p> | ||
+ | <p>- PCR clean up</p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p>BBa_K929000 & BBa_J52034 </p> | ||
+ | <ul> | ||
+ | <blockquote> | ||
+ | <p>- Inoculation </p> | ||
+ | </blockquote> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p><strong>August 9</strong></p> | ||
+ | <p>BBa_K929000 & BBa_J52034 </p> | ||
+ | <p>- Plasmid extraction: J52034 is not transformed</p> | ||
+ | <p>MLS.pSB1C3 & GFP.PolyA</p> | ||
+ | <p>- Ligation</p> | ||
+ | <p>- Transformation to SURE on CAM plate</p> | ||
+ | <p> </p> | ||
+ | |||
</div> | </div> | ||
</div> | </div> |
Revision as of 17:01, 19 September 2013
June 2013
· Inoculation of pBlueScript KS(+) for training
· Autoclave basic materials
· Preparing LB
· LB-Ampicillin plates poured
Dry lab
Protocols review
August 2013
August 5
GFP (25)PCR Check with VR VF2
- Gel check(overan)
BBa_J176171
- Digestion using SpeI and PstI-HF to extract the backbone
PolyA / BBa_K404108
- Digestion using XbaI and PstI-HF
- Gel electrophoresis
- Run Gel
Inoculation of pSB1C3
August 6
GFP (25)
- Repeat PCR check
- Run Gel
pSB1C3
- Plasmid extraction
PolyA / BBa_K404108
- Gel purification
- Gel check
BBa_J176171
- PCR clean up
BBa_K929000 & BBa_J52034
- Transformation using CAM plate
MLS
- PCR amplify MLS in RFC 10 and 25 format
August 7
MLS (RFC 10 & 25)
- PCR clean up
- Nanodrop for the PCR clean up product
- Gel check for the PCR clean up product
- Digestion using EcoRI and PstI
PolyA / BBa_K404108
- Gel check
pSB1C3
- Digest with EcoRI & PstI
- Run gel and cut gel
GFP(25)
- Digestion with EcoRI & SpeI
- Run gel and cut gel
August 8
GFP25 & pSB1C3
- Gel purification
- Nanodrop
- Gel check
polyA
- Digestion with EcoRI and XbaI
- PCR clean up
BBa_K929000 & BBa_J52034
- Inoculation
August 9
BBa_K929000 & BBa_J52034
- Plasmid extraction: J52034 is not transformed
MLS.pSB1C3 & GFP.PolyA
- Ligation
- Transformation to SURE on CAM plate