Template:Kyoto/Notebook/Aug 17

From 2013.igem.org

(Difference between revisions)
(Liquid Culture)
(Restriction Enzyme Digestion)
Line 89: Line 89:
*at 37°C, for 1h
*at 37°C, for 1h
{| class="wikitable"
{| class="wikitable"
-
! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
+
! ||8/17 K748002(lysis3) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||2.0µL||1.0µL||1.0µL||3.0µL||3.0µL||20µL||30µL
 +
|-
 +
|1 cut||1.0µL||0.2µL||--||1.0µL||1.0µL||6.8µL||10µL
 +
|-
 +
|1 cut||1.0µL||--||0.2µL||1.0µL||1.0µL||6.8µL||10µL
 +
|-
 +
|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL
 +
|}
 +
*at 37°C, for 1h
 +
{| class="wikitable"
 +
! ||8/17 I13504(RBS-GFP-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
|-
|-
|2 cuts||2.0µL||1.0µL||1.0µL||3.0µL||3.0µL||20µL||30µL
|2 cuts||2.0µL||1.0µL||1.0µL||3.0µL||3.0µL||20µL||30µL

Revision as of 15:53, 23 September 2013

Contents

Aug 17

Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles

File:Image

Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Image

Liquid Culture

Nakamoto

Samplemedium
8/15 BBa_K112806(lysis2) -(1)Plusgrow medium(+CP)
8/15 BBa_Q04400(tetR) -(1)Plusgrow medium(+Amp)

Restriction Enzyme Digestion

Murata,Murakami,Nakamoto,Ashida,Matsumoto and Yoshida

8/11 J23100 -(2)SPeIPstIBufferMilliQtotal
2 cuts4.0µL1.0µL1.0µL3.0µL21µL30µL
1 cut1.3µL0.2µL--1.0µL7.5µL10µL
1 cut1.3µL--0.2µL1.0µL7.5µL10µL
NC1.3µL----1.0µL7.7µL10µL
  • at 37°C, for 1h
8/15 B0034(RBS) -(2)SPeIPstIBufferMilliQtotal
2 cuts6.0µL1.0µL1.0µL3.0µL19µL30µL
1 cut2.0µL0.2µL--1.0µL6.8µL10µL
1 cut2.0µL--0.2µL1.0µL6.8µL10µL
NC2.0µL----1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K117000(lysis1) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K748002(lysis3) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 I13504(RBS-GFP-DT) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h

Master Plate

Nakamoto

NumberUse LB plate (+CP)
1BBa_K112806(lysis2) -(1)
NumberUse LB plate (+Kana)
1BBa_Q04400(tetR) -(1)