"
Team:TU-Munich/Notebook/Labjournal
From 2013.igem.org
// put the footer in the right place
$("#footer-box").prepend($("#social-footer"));
// implement image preloading
var images = new Array()
function preload() {
for (i = 0; i < preload.arguments.length; i++) {
images[i] = new Image()
images[i].src = preload.arguments[i]
}
}
// preload menu backgrounds
preload( "
",
"", "
", "
", "
", "
" );
// preload team pictures
if ( $("div#teamfield").length > 0 ) {
preload( "
",
"", "
", "
", "
", "
", "
", "
", "
", "
", // Katrin "
", "
", "
", "
", "
", "
", "
", "
", "
", // Rosario "
", "
", "
", "
", "
", "
", "
", "
", "
", // Fabian "
", "
", "
", "
", "
", "
", "
", "
", "
", // Andreas "
", "
", "
", "
", "
", "
", "
", "
", "
", // Louise "
", "
", "
", "
", "
", "
", "
", "
", "
", // Johanna "
", "
", "
", "
", "
", "
", "
", "
", "
", // Meike "
", "
", "
", "
", "
", "
", "
", "
", "
", // Volker "
", "
", "
", "
", "
", "
", "
", "
", "
", // Polte "
", "
", "
", "
", "
", "
", "
", "
", "
", // Leonie "
", "
", "
", "
", "
", "
", "
", "
", "
", // Philipp "
", "
", "
", "
", "
", "
", "
", "
", "
", // Jeff "
", "
", "
", "
", "
", "
", "
", "
", "
", // Chris "
", "
", "
", "
", "
", "
", "
", "
", "
", // Flo "
", "
", "
", "
", "
", "
", "
", "
" );
}
// Slideshows
$('.bxslider').bxSlider({
responsive: false, auto: true, autoHover: true
});
$('.bxgallery').bxSlider({
slideMargin: 10, minSlides: 3, maxSlides: 3, moveSlides: 1, slideWidth: 5000
});
$("ul.bxgallery img").slimbox({ loop: true }, function(el) { url = el.src; url = url.substring(0, url.lastIndexOf('/')); url = url.replace('/thumb/', '/'); // description = $(el).parents("div.thumbinner").children("div.thumbcaption").text(); return [url, ]; }, function(el) { return (this == el) || (this.parentNode.parentNode && (this.parentNode.parentNode == el.parentNode.parentNode)); });
// Counter and Countdown
function render_counter(c) { i = 0; iid = window.setInterval(function(){ if ( (c-i) > (c/200) ) { $('span#counter').html(i); i += Math.round(c/200); } else { $('span#counter').html(c); window.clearInterval(iid); } }, 10); }
if ($('span#counter').length > 0) { $.ajax({ url: "https://2013.igem.org/Special:PopularPages", success: function( html ) { dom = $.parseHTML(html); visitors = $(dom).find('a[title="Team:TU-Munich"]').parent().text(); visitors = visitors.substring(visitors.indexOf('(')+1); visitors = visitors.substring(0, visitors.indexOf(' ')); visitors = visitors.replace(',', ); render_counter(visitors); }, error: function( xhr, status ) { render_counter(4700); } }); }
if ($('span#countdown) { clock = window.setInterval(function(){ jetzt = new Date(); time_left = Date.UTC(2013, 9, 5, 4, 0, 0) - Date.UTC(jetzt.getUTCFullYear(), jetzt.getUTCMonth(), jetzt.getUTCDate(), jetzt.getUTCHours(), jetzt.getUTCMinutes(), jetzt.getUTCSeconds()); left_sec = (time_left/1000)%60; left_sec = (left_sec < 10) ? "0" + left_sec : left_sec; left_min = Math.floor(time_left/60000)%60; left_min = (left_min < 10) ? "0" + left_min : left_min; left_h = Math.floor(time_left/3600000)%24; left_h = (left_h < 10) ? "0" + left_h : left_h; left_d = Math.floor(time_left/86400000); left_d = (left_d == 1) ? left_d + " day" : left_d + " days"; $('span#countdown').html(left_d + " " + left_h + ":" + left_min + ":" + left_sec); }, 1000); }
// Animate teamfield
if ( $("div#teamfield").length > 0 ) {
var $members = $("div#teamfield a");
$("body").mousemove(function(event){ for (i=0; i<$members.length; i++) {
if ( $members.eq(i).offset().left > event.pageX ) {
if ( $members.eq(i).offset().top > event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("top-left");
} else if ( $members.eq(i).offset().top <= event.pageY && ( $members.eq(i).offset().top + $members.eq(i).height() ) >= event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("left");
} else if ( ( $members.eq(i).offset().top + $members.eq(i).height() ) < event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("bottom-left");
}
} else if ( $members.eq(i).offset().left <= event.pageX && ( $members.eq(i).offset().left + $members.eq(i).width() ) >= event.pageX ) {
if ( $members.eq(i).offset().top > event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("top");
} else if ( $members.eq(i).offset().top <= event.pageY && ( $members.eq(i).offset().top + $members.eq(i).height() ) >= event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("front");
} else if ( ( $members.eq(i).offset().top + $members.eq(i).height() ) < event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("bottom");
}
} else if ( ( $members.eq(i).offset().left + $members.eq(i).width() ) < event.pageX ) {
if ( $members.eq(i).offset().top > event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("top-right");
} else if ( $members.eq(i).offset().top <= event.pageY && ( $members.eq(i).offset().top + $members.eq(i).height() ) >= event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("right");
} else if ( ( $members.eq(i).offset().top + $members.eq(i).height() ) < event.pageY ) {
$members.eq(i).removeClass();
$members.eq(i).addClass("bottom-right");
}
}
} });
}
});
100 bp GeneRuler:
PageRuler Plus:
Labjournal
Week 1
Monday, April 22nd
Transformation of E. coli XL1 blue with Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3)
Investigator: Jeff, Rosario
Aim of the experiment: Transformation of Phytochrome B for protein fusion.
Procedure:
- CaCl2 competent E. coli XL1-Blue cells were put out from the stock in -80 °C freezer and were gently thawed on ice.
- 2 µl of DNA was added to 100 µl of competent cells and gently mixed.
- 30 min incubation on ice
- 5 min. heat shock at 37 °C
- Adding of 1 ml LB-medium to each tube.
- Incubation for 45 min at 37 °C in the 180 rpm cell-culture shaker.
- 100 µl of the cell suspension was plated on one chloramphenicol plate.
- The rest were centrifuged for 1 min at 13000 rpm and the supernatant was dicarded.
- The pellet was resuspended in 100 µl of LB-medium and this concentrated cell suspension was plated again on a new chlorampenicol plate.
Miniprep of pTUM100 with pGAL, pTEF1, pTEF2, pADH and RFC25 compatible RFP generator
Investigator: Jeff, Rosario
Aim of the experiment: Miniprep of pTUM100 with pGAL, pTEF1, pTEF2, pADH and RFC25 compatible RFP generator
Procedure:
- Miniprep was performed after manufacturer's protocol (QIAprep Miniprep, QIAGEN)
Sequencing of RFP-Generator (RFC25, pSB1C3)
Investigator: Jeff, Rosario
Aim of the experiment: Sequencing of RFP-Generator (RFC25, pSB1C3)
Procedure:
Sequencing batch were prepared after manufacturer's protocol. (15 µl of plasmid DNA (50 - 100 ng) and 2 µl sequencing primer)
Tuesday, April 23rd
Picking of of E. coli XL1 blue with Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3)
Investigator: Jeff, Rosario, Florian
Aim of the experiment: Picking of of E. coli XL1 blue with Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3)
Procedure:
- pSB1C3 plasmid with BBa_K801031 (PhyB 2 - 908 aa, RFC25): Colonies were picked from chloramphenicol plates.
- Picked pipette tips was transferred into cell-culture tubes with air-permeable, sterile cover. Each tube contain 4 mL of LB-medium + 4 µL chloramphenicol(1000x).
- 4 colonies were picked.
- These tubes were transferred in a cell culture shaker at 37 °C and were incubated overnight
Analytical digestion and gelelectrophoresis of RFP-generator (RFC25, pSB1C3, P4 & P5)
Investigator: Jeff, Rosario, Florian
Aim of the experiment: Analytical digestion and gelelectrophoresis of RFP-generator (RFC25, pSB1C3, P4 & P5).
Procedure:
- Batch for analytical digestion for P4 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P4 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | NgoMIV (10 U/µl) |
| 0.25 µl | AgeI-HF (20 U/µl) |
| 15 µl | ddH2O |
| =20 µl | TOTAL |
- Batch for analytical digestion for P5 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P5 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | NgoMIV (10 U/µl) |
| 0.25 µl | AgeI-HF (20 U/µl) |
| 15 µl | ddH2O |
| =20 µl | TOTAL |
- Incubation for 90 min at 37 °C.
- Analytical gelelectrophoresis was performed at 90 V for 60 min.
Results:
| 1 kbp ladder DNA ladder | P4 | P5 |
| Mutation successful | Mutation successful! |
- Parts are compliant and do not contain RFC25 forbidden restriction sites.
Sequencing of pTUM vectors with pGAL, pADH, pTEF1, pTEF2
Investigator: Jeff, Rosario, Florian
Aim of the experiment: Sequencing of pTUM vectors with pGAL, pADH, pTEF1, pTEF2
Procedure:
Sequencing batch were prepared after manufacturer's protocol. (15 µl of plasmid DNA (50 - 100 ng) and 2 µl sequencing primer).
The different vectors we sequenced received the following barcodes:
- ADH in pTUM100: FR01002265
- TEF1 in pTUM100: FR01002266
- TEF2 in pTUM100: FR01002266
- GAL in pTUM100: FR01002268
Sequencing of TEF2 in pTUM100 was not interpretable. The other sequences were consistent with the sequences in the parts registry.
Wednesday, April 24th
Miniprep of Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3)
Investigator: Jeff, Florian
Aim of the experiment: Miniprep of Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3).
Procedure:
- Miniprep was performed after manufacturer's protocol (QIAprep Miniprep, QIAGEN)
Analytical digestion and gelelectrophoresis of Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3), P7 - P10
Investigator: Jeff, Florian
Aim of the experiment: Analytical digestion and gelelectrophoresis of Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3), P7 - P10.
Procedure:
- Batch for analytical digestion for P7 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P7 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | NgoMIV (10 U/µl) |
| 0.25 µl | AgeI-HF (20 U/µl) |
| 15 µl | ddH2O |
| =20 µl | TOTAL |
- Batch for analytical digestion for P8 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P8 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | NgoMIV (10 U/µl) |
| 0.25 µl | AgeI-HF (20 U/µl) |
| 15 µl | ddH2O |
| =20 µl | TOTAL |
- Batch for analytical digestion for P9 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P9 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | NgoMIV (10 U/µl) |
| 0.25 µl | AgeI-HF (20 U/µl) |
| 15 µl | ddH2O |
| =20 µl | TOTAL |
- Batch for analytical digestion for P10 with NgoMIV+AgeI-HF
| volume | reagent |
| 2.5 µl | Plasmid DNA P10 |
| 2 µl | NEBuffer 4 (10x) |
| 0.25 µl | Ngo |
