Team:SDU-Denmark/labnotes7

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= Lab notes (16/9 - 22/9) =
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= Lab notes (23/8 - 29/8) =
__TOC__
__TOC__
 +
 +
== Photosensor ==
 +
=== Insertion of B0015 in pSB3C5 and pSB3T5 ===
 +
 +
'''Date:''' 8/23 - 8/29 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:''' [https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#DE1.3 DE1.3][https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1 RD1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#LG1.2 LG1.2][https://2010.igem.org/Team:SDU-Denmark/protocols#CC1.1 CC1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#TR1.1 TR1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.3 CP1.3]<Br>
 +
==== Pfu PCR amplification and purification of B0015 ====
 +
'''Date:''' 8/23 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1]<br>
 +
'''Notes:'''<br>
 +
4 PCR reactions are prepared. 2uL Miniprep of pSB1AK3 (white 43) are distrubuted in each tube. PCR tubes are marked B00153.A-D.<br>
 +
Premix x5:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>pfu buffer + MgSO<small>4</small></td>
 +
      <td>25uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>dNTP mix</td>
 +
      <td>7.5uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>VF2 primer</td>
 +
      <td>7.5uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>VR primer</td>
 +
      <td>7.5uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>H<small>2</small>0</td>
 +
      <td>190uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pfu polymerase&nbsp;</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
</table><br><br>
 +
PCR program:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>start</td>
 +
      <td>94C</td>
 +
      <td>3min</td>
 +
    </tr>
 +
    <tr>
 +
      <td>denaturating</td>
 +
      <td>94C</td>
 +
      <td>2min</td>
 +
    </tr>
 +
    <tr>
 +
      <td>annealing</td>
 +
      <td>55C</td>
 +
      <td>30s</td>
 +
    </tr>
 +
    <tr>
 +
      <td>elongation</td>
 +
      <td>72C</td>
 +
      <td>30s</td>
 +
    </tr>
 +
    <tr>
 +
      <td>go to</td>
 +
      <td>2</td>
 +
      <td>rep.29x</td>
 +
    </tr>
 +
    <tr>
 +
      <td>end</td>
 +
      <td>72C</td>
 +
      <td>5min</td>
 +
    </tr>
 +
    <tr>
 +
      <td>hold</td>
 +
      <td>4C</td>
 +
      <td></td>
 +
    </tr>
 +
</table><br>
 +
5uL of the PCR product was loaded onto a 2% agarose gel. Gene ruler 100bp DNA ladder was used ad marker.<br><br>
 +
'''Results:'''<br>
 +
'''Analysis:'''<br>
 +
A strong band was observed at app. 450bp, and the rest of the PCR product was purified according to protocol using the GFX purification kit.<br>
 +
End conc.: 48ng/uL <br><br>
 +
 +
==== Restriction digest of B0015 ====
 +
'''Date:''' 8/24 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1 RD1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#DE1.3 DE1.3]<br>
 +
'''Notes:'''<br>
 +
Restriction mixture B0015:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>H<small>2</small>O</td>
 +
      <td>38uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>FD green buffer</td>
 +
      <td>8uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>EcoRI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PstI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>B0015</td>
 +
      <td>30uL</td>
 +
    </tr>
 +
</table><br><br>
 +
 +
The digested sample was loaded onto a 2% agarose extraction gel. Uncut B0015 was used as controle. Gene ruler 100bp DNA ladder was used as marker.<br>
 +
DNA was extracted from gel according to protocol.<br><br>
 +
'''Results:'''<br>
 +
DNA conc:
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>sample</td>
 +
      <td>conc. (ng/uL)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>B0015</td>
 +
      <td>6.5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB3C5</td>
 +
      <td>50.8</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB3T5</td>
 +
      <td>9.9</td>
 +
    </tr>
 +
</table><br><br>
 +
'''Analysis:'''
 +
the purified DNA was used for ligation.<br><br>
 +
 +
==== Ligation of PS and pSB1C3 and pCB1AK3 ====
 +
'''Date:''' 8/24 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#LG1.2 LG1.2]<br>
 +
'''Notes:'''<br>
 +
For each of the ligations three ligation mixtures were prepared. vector concentrations of 10n0g/uL (pSB3C5) and 50ng/uL (pSB3T5) respectively was used for each mixture. Appropiate amount of insert was added to reach vector:insert ratios of 1:1, 1:3 and 1:6 respectively. <br>
 +
Ligation mixtures (B0015 in pSB3C5):<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td></td>
 +
      <td>L1</td>
 +
      <td>L2</td>
 +
      <td>L3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>T4 ligase buffer</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>T4 ligase</td>
 +
      <td>1uL</td>
 +
      <td>1uL</td>
 +
      <td>1uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB3C5</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>B0015 </td>
 +
      <td>0.7uL</td>
 +
      <td>2uL</td>
 +
      <td>4.5uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>H<small>2</small>0</td>
 +
      <td>14.3uL</td>
 +
      <td>13uL</td>
 +
      <td>10.5uL</td>
 +
    </tr>
 +
</table><br>
 +
Ligation mixtures (B0015 in pSB3T5):<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td></td>
 +
      <td>L1</td>
 +
      <td>L2</td>
 +
      <td>L3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>T4 ligase buffer</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>T4 ligase</td>
 +
      <td>1uL</td>
 +
      <td>1uL</td>
 +
      <td>1uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB3T5</td>
 +
      <td>5uL</td>
 +
      <td>5uL</td>
 +
      <td>5uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>B0015 </td>
 +
      <td>0.5uL</td>
 +
      <td>1uL</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>H<small>2</small>0</td>
 +
      <td>11.5uL</td>
 +
      <td>11uL</td>
 +
      <td>10uL</td>
 +
    </tr>
 +
</table><br><br>
 +
The samples was incubated at 17C ON at used for transformation<br><br>
 +
 +
==== Transfomation of ligated plasmid in Top 10 E.coli ====
 +
 +
'''Date:''' 8/25 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#CC1.1 CC1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#TR1.1 TR1.1]<br>
 +
'''Notes:'''<br>
 +
The compotent cells and transformation was carried out according to protocol.<br><br>
 +
'''Results:'''<br>
 +
the controle plates were okay, and there were many colonies on plates with cells transformed with either of the ligation mixtures..<br><br>
 +
'''Analysis:'''<br>
 +
The transformation was successfull and colonies was selected and used in coloni PCR.<br>
 +
--[[User:Tipi|Tipi]] 13:41, 26 September 2010 (UTC)<br><br>
 +
=== Colony PCR on B0017 ===
 +
Date: 27/8<br>
 +
Done by: LC<br>
 +
Methods: PCR<br>
 +
Protocols: [https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.3 CP1.3]<br>
 +
Notes: <br>
 +
Premix:<br>
 +
12,5 µl 10xTAQ Buffer<br>
 +
5 µl MgCl2 <br>
 +
5 µl VF2 <br>
 +
5 µl VR <br>
 +
2,5 µl dNTP<br>
 +
17,5 µl H2O<br>
 +
5/8 µl TAQ Polymerase<br>
 +
<br>
 +
9,5 µl Premix were added to 15 µl of H2O containing the lysed cells.<br>
 +
<br>
 +
PCR Program:<br>
 +
<table style="text-align: left;" border="1" cellpadding="2"
 +
cellspacing="2">
 +
<tr>
 +
<td style="vertical-align: top;">Start<br>
 +
</td>
 +
<td style="vertical-align: top;">94&nbsp; C<br>
 +
</td>
 +
<td style="vertical-align: top;">2 min<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">Denaturing<br>
 +
</td>
 +
<td style="vertical-align: top;">94 C<br>
 +
</td>
 +
<td style="vertical-align: top;">1 min<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">Annealing<br>
 +
</td>
 +
<td style="vertical-align: top;">55 C<br>
 +
</td>
 +
<td style="vertical-align: top;">1 min<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">Elongation<br>
 +
</td>
 +
<td style="vertical-align: top;">72 C<br>
 +
</td>
 +
<td style="vertical-align: top;">30 sec<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">Goto2<br>
 +
</td>
 +
<td style="vertical-align: top;">rep<br>
 +
</td>
 +
<td style="vertical-align: top;">29x<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">End<br>
 +
</td>
 +
<td style="vertical-align: top;">72 C<br>
 +
</td>
 +
<td style="vertical-align: top;">3 min<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">Hold<br>
 +
</td>
 +
<td style="vertical-align: top;">4 C<br>
 +
</td>
 +
<td style="vertical-align: top;"><br>
 +
</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
Results: <br>
 +
[[Image:Team-SDU-Denmark-cPCRB0017.jpg|300px]] <br>
 +
No useable results, only unclear bands around 120 BP, which seem to be the result of mispriming with VR on B0010.
 +
 +
== Retinal ==
 +
 +
==== Futher PCR on POT2 with NinaB (New Primers NO. 5) ====
 +
Date: 23/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: PCR<br>
 +
protocos:[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1]
 +
Notes:<br>
 +
NinB2fw and NinaB2rv was used.<br>
 +
PCR were run programed as:<br>
 +
<html>
 +
<head>
 +
  <meta content="text/html; charset=ISO-8859-1"
 +
http-equiv="content-type">
 +
  <title></title>
 +
</head>
 +
<body>
 +
<table style="text-align: left; width: 300px;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>PCR</td>
 +
      <td>Temp. (C)</td>
 +
      <td>Time (min)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Start</td>
 +
      <td>95</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Denaturing</td>
 +
      <td>95</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Anneling</td>
 +
      <td>67,9</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Elongation</td>
 +
      <td>72</td>
 +
      <td>4</td>
 +
    </tr>
 +
    <tr>
 +
      <td>End</td>
 +
      <td>72</td>
 +
      <td>5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Hold</td>
 +
      <td>4</td>
 +
      <td>indef.</td>
 +
    </tr>
 +
</table>
 +
<br>
 +
</body>
 +
</html>
 +
<br>
 +
PCR product from gradient PCR (d. 20/8-10), tube no. 5, was used as template.<br>
 +
The other tubes were pooled.<br>
 +
 +
=== PCR on POT2 with NinaB (New Primers) ===
 +
Start date: 24/8<br>
 +
Methods: Ligation, Competent cells, Transformation<br>
 +
Protocols: [https://2010.igem.org/Team:SDU-Denmark/protocols#LG1.1 LG1.1], [https://2010.igem.org/Team:SDU-Denmark/protocols#CC1.1 CC1.1], [https://2010.igem.org/Team:SDU-Denmark/protocols#TR1.1 TR1.1]
 +
 +
==== DNA purification from PCR ====
 +
Date: 20/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: DNA purification from PCR<br>
 +
protocos:GFX purification from PCR - kit
 +
<br><br>
 +
One of the pooled tubes was eluted in 200µL, the others was eluted in 20µL<br>
 +
200µL nanodrop: 16,4 ng/µL<br>
 +
20µL nanodrop: 133,9ng/µL<br>
 +
 +
==== Restriction Digest ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: Restriction Digest<br>
 +
protocos:RD1.1[https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1]
 +
Notes:<br>
 +
Restriction mixture:<br>
 +
<html>
 +
<head>
 +
  <meta content="text/html; charset=ISO-8859-1"
 +
http-equiv="content-type">
 +
  <title></title>
 +
</head>
 +
<body>
 +
<table style="text-align: left; width: 100px;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>38 µL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>8 µL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>4&nbsp;µL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>4&nbsp;µL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>30 µL</td>
 +
    </tr>
 +
</table>
 +
<br>
 +
</body>
 +
</html>
 +
Gel was run with uncut controles:<br>
 +
 +
==== Gel purification ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: gel purifikation<br>
 +
protocos:GFX gel purifikation kit
 +
Notes:<br>
 +
Purifide products was Nanodroped:<br>
 +
NinaB 1: 4,5 ng/µL<br>
 +
NinaB 2: 1,15 ng/µL<br>
 +
NinaB 3: 7,74 ng/µL<br>
 +
PSB1C3: 25,66 ng/µL<br>
 +
Nina B pooled: 4,5 ng/µL<br>
 +
 +
==== Ligation ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: Ligation<br>
 +
protocos:[https://2010.igem.org/Team:SDU-Denmark/protocols#LG1.3 L1.3]
 +
Notes:<br>
 +
3 ligatons mixtures was made:<br>
 +
1:1 volumens 1 plasmid:5 insert
 +
1:3 volumens 1 plasmid:15 insert
 +
1:6 volumens 1 plasmid:30 insert
 +
 +
==== Colony PCR on ligation from 24/8 ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: Colony PCR<br>
 +
protocos:[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1]
 +
Notes:<br>
 +
15 colonies were picked form different plates, with differnt plasmid to insert ratio: colonie's 1,2,3,13 were picked from plates with 1:1 plasmid to insert ratio, colonie's 4,5,6 were picked from plates with 1:3 plasmid to insert ratio and colonie's 7,8,9,10,11,12,14,15 were picked from plates with 1:6 plasmid to insert ratio.<br>
 +
The PCR program was:<br>
 +
<html>
 +
<head>
 +
  <meta content="text/html; charset=ISO-8859-1"
 +
http-equiv="content-type">
 +
  <title></title>
 +
</head>
 +
<body>
 +
<table style="text-align: left; width: 300px;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>PCR</td>
 +
      <td>Temp. (C)</td>
 +
      <td>Time (min)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Start</td>
 +
      <td>95</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Denaturing</td>
 +
      <td>95</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Anneling</td>
 +
      <td>68,0</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Elongation</td>
 +
      <td>72</td>
 +
      <td>4</td>
 +
    </tr>
 +
    <tr>
 +
      <td>End</td>
 +
      <td>72</td>
 +
      <td>5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Hold</td>
 +
      <td>4</td>
 +
      <td>indef.</td>
 +
    </tr>
 +
</table>
 +
<br>
 +
</body>
 +
</html>
 +
A gel was run on the PCR products:<br>
 +
Futher experiments and PCR was run on tubes: 5,6,10,11 because they have the greatest yeild.<br>
 +
 +
==== Restriction Digest ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: Restriction Digest<br>
 +
protocos:[https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1 RD1.1]
 +
Notes:<br>
 +
Restriction digest was performed on tubes 5,6,10 and 11 to test for insertion of ninaB (Correct orientation) XbaI and SPEI was used and a gel was run:<br>
 +
 +
==== Colony PCR on ligation from 24/8 (colonies 5,6,10 and 11 + new colonies) ====
 +
Date: 24/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: Colony PCR<br>
 +
protocos:[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1]
 +
Notes:<br>
 +
8 new colonies (16-23) were chosen in addition to colonies 5,6,10 and 11 form 25I8-10 colonie PCR.<br>
 +
PCR were run with TAQ polymerase and VF2 + VR primers according to the following program:<br>
 +
<html>
 +
<head>
 +
  <meta content="text/html; charset=ISO-8859-1"
 +
http-equiv="content-type">
 +
  <title></title>
 +
</head>
 +
<body>
 +
<table style="text-align: left; width: 300px;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>PCR</td>
 +
      <td>Temp. (C)</td>
 +
      <td>Time (min)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Start</td>
 +
      <td>94</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Denaturing</td>
 +
      <td>94</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Anneling</td>
 +
      <td>55,0</td>
 +
      <td>0.5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Elongation</td>
 +
      <td>72</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>End</td>
 +
      <td>72</td>
 +
      <td>5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Hold</td>
 +
      <td>4</td>
 +
      <td>indef.</td>
 +
    </tr>
 +
</table>
 +
A gel was run on the products:<br>
 +
==== PCR on NinaB with old primers (NinaBfw and NinaBrv) ====
 +
Date: 27/8<br>
 +
Done by: Marie & Tommy<br>
 +
Methods: PCR<br>
 +
protocos:CP1.1[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1]
 +
Notes:<br>
 +
PCR was run on NinaB (from PCR with NinaBfw and NinaBrv), this time with the old primers (NinaBfw and NinaBrv) in an attempt to add the E and P restriction sites to the NinaB fracment.<br>
 +
The PCR program was set to:<br>
 +
<html>
 +
<head>
 +
  <meta content="text/html; charset=ISO-8859-1"
 +
http-equiv="content-type">
 +
  <title></title>
 +
</head>
 +
<body>
 +
<table style="text-align: left; width: 300px;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>PCR</td>
 +
      <td>Temp. (C)</td>
 +
      <td>Time (min)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Start</td>
 +
      <td>95</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Denaturing</td>
 +
      <td>95</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Anneling</td>
 +
      <td>55,0</td>
 +
      <td>0.75</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Elongation</td>
 +
      <td>72</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Denaturing</td>
 +
      <td>94</td>
 +
      <td>1</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Anneling</td>
 +
      <td>73,0</td>
 +
      <td>0.75</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Elongation</td>
 +
      <td>72</td>
 +
      <td>2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>End</td>
 +
      <td>72</td>
 +
      <td>5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Hold</td>
 +
      <td>4</td>
 +
      <td>indef.</td>
 +
    </tr>
 +
</table>
 +
NinaB From second new primer PCR (23/8, purifid 24/8) was used as templated.<br>

Latest revision as of 22:31, 23 October 2010