Team:NJU China/Notebook

From 2013.igem.org

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13th: We analyzed the data.</br>
13th: We analyzed the data.</br>
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                    <h2 id="July">July</h2>
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                    <div class="ss-right">
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                    <h2>2013</h2>
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<h3>
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<a>WEEK 8</a>
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<span>19th-24th, June</span>
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<h3>
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<a>siRNA screening (failed)</a>
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<span>
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8th: We cultured Hep G2 cells in 2 12-well plates for 24-hour cell collection and 48-hour cell collection.</br>
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9th: We transfected cells with empty Lipo, HbsAg plasmids, HbsAg plasmids and 467 siRNA plasmids, HbsAg plasmids and 516 siRNA plasmids, respectively.</br>
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10th: We collected cells 24 hours after transfection, and preserved them in Trizol.</br>
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11th: We collected 48 hours after transfection, and then extracted RNA from 2 groups of cells (24 hours and 48 hours).</br>
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12th: We did RT-PCR and qPCR with all RNA samples.</br>
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13th: We analyzed the data.</br>
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</span>   
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<a>　　Absolute quantification of exosomes (failed)</a>
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<span>
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5th: We cultured 293t cells.</br>
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6th: Cells we cultured on 5th July were contaminated by bacteria.</br>
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7th: We subcultured another cell line of 293t cells.</br>
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9th: We cultured 293t cells in 6 flasks.</br>
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10th:We transferred 293T cells with plasmids.</br>
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11th: We collected exosomes 24 hours after transfection.</br>
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12th: A part of cells died, failed to collect 48-hour exosomes.</br>
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13th: We examined protein concentration of 24-hour exosomes, started to extract RNA from 24h-cells and 24h-exosomes and preserved the rough RNA extract solution with isopropyl alcohol at 4℃ overnight.</br>
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14th：We continued finish RNA extraction, then did RT-PCR and qPCR.</br>
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15th: We analyzed data.</br>
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</span>     
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<a>　　Luciferase Assay</a>
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<span>
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11th: Construction of plasmids: obtain vector and segment.</br>
 +
12th: We combined vector and segment with T4 ligase, then transformed the recombined plasmids into E.coli DH5α competent cells and spread them on solid LB culture plates by streak plate method.</br>
 +
13th: No single bacterial colony was found.</br>
 +
14th: No single bacterial colony was found again, redid previous steps(obtain the target segments, combined vector and segment, and then transferred it into E.coli )</br>
 +
15th: Single bacterial colonies were found. We picked single colonies and transferred them into liquid LB culture medium with ampicillin, and shaken overnight at 37℃.</br>
 +
16th: The transformed E.coli cells failed to reproduce in LB culture medium.</br>
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</span> 
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<a>Others</a>
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<span>
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5th: We thawed 293t cells.</br>
 +
6th: We subcultured HepG2 cells.</br>
 +
7th: E.coli cells containing HBSag overexpressed plasmids were transferred respectively into LB culture medium with ampicillin, and shaken overnight at 37℃.</br>
 +
8th: We extracted HBSag overexpressed plasmids from E.coli cells.</br>
 +
11th: We thawed 293t cells and subcultured HepG2 cells. E.coli cells containing GFP plasmids were transferred into LB culture medium with ampicillin, and shaken overnight at 37℃.</br>
 +
12th: We extracted GFP plasmids from E.coli cells.</br>
 +
14th: We thawed HepG2 cells and 293T cells.</br>
 +
15th: We thawed 293T cells.</br>
 +
</span>                   
</h3>
</h3>
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Revision as of 19:21, 26 September 2013

 

Notebook

March

2013

WEEK 1 25th-28th, March

Experiment training We learnt to culture the 293t cells.
We learnt to extract plasmids and transfect it into 293t cells.
We learnt to collect exosomes.
We learnt to extract RNA from exosomes and cells.
We learnt to do RT-PCR and qPCR.

April

2013

WEEK 2 14th-18th, April

siRNA screening (Failed) 14th: We extracted plasmids and cultured the 293t cells.
15th: We transfected plasmids into 293t cells.
16th: We collected cells and preserved it in Trizol.
17th: We extracted RNA, then did RT-PCR with the RNA.
18th: We did qPCR with the cDNA we got on 17th April.
21th: We reexamined the concentration of RNA, and redid qPCR.

WEEK 3 27th, April – 1st, May

siRNA screening (Failed) 27th: We cultured the 293t cells in a 12-well plate.
28th: We transfected 293t cells.
29th: We collected cells and extracted RNA from it.
30th: We did RT-qPCR with the RNA we extracted on 29th April.
1st: We did qPCR with the cDNA we got on 30th April.

May

2013

WEEK 4 8th-13th, May

siRNA screening (Failed), examination whether siRNA are capsulated into exosomes (Success) 8th: We extracted plasmids of 3 kinds of siRNA.
9th: We cultured 293t cells in eight D10 dishes and a 12-well plate.
10th: We extracted 2 kinds of over-expression plasmids and examined the concentration of it. Then we transfect these plasmids into 293t cells respectively.
11th: We collected cells and exosomes from 8 D10 dish, and cells from 12-well plate.
12th: We extracted RNA from cells and exosomes.
13th: We did RT-PCR and qPCR.

WEEK 5 15th-18th, May

　　siRNA screening (Failed) 15th: We cultured the 293t cells in a 12-well plate.
16th: We transfected 293t cells.
17th: We collected cells and extracted RNA from then.
18th: We did RT-PCR and qPCR.

WEEK 6 27th-31th, May

　　siRNA screening (Failed) 27th: We cultured the 293t cells in 2 12-well plates.
28th: We transfected 293t cells, with lipofectamine 2000.
29th: We collected cells and extracted RNA from them.
30th: We continued to extract RNA, and examine the concentration of total RNA. Then we did RT-PCR.
31st : We did qPCR.

June

2013

WEEK 7 19th-24th, June

siRNA screening (eliminated 308 siRNA, 467 siRNA and 516 siRNA seem to have good effect) 19th: We cultured 293t cells in 12-well plates.
20th: We transfected 293t cells.
21st: We collected cells and preserved in Trizol.
22nd: We extracted RNA from cells and did RT-PCR.
24th: We did qPCR.

July

2013

WEEK 8 19th-24th, June

siRNA screening (failed) 8th: We cultured Hep G2 cells in 2 12-well plates for 24-hour cell collection and 48-hour cell collection.
9th: We transfected cells with empty Lipo, HbsAg plasmids, HbsAg plasmids and 467 siRNA plasmids, HbsAg plasmids and 516 siRNA plasmids, respectively.
10th: We collected cells 24 hours after transfection, and preserved them in Trizol.
11th: We collected 48 hours after transfection, and then extracted RNA from 2 groups of cells (24 hours and 48 hours).
12th: We did RT-PCR and qPCR with all RNA samples.
13th: We analyzed the data.

July

2013

WEEK 8 19th-24th, June

siRNA screening (failed) 8th: We cultured Hep G2 cells in 2 12-well plates for 24-hour cell collection and 48-hour cell collection.
9th: We transfected cells with empty Lipo, HbsAg plasmids, HbsAg plasmids and 467 siRNA plasmids, HbsAg plasmids and 516 siRNA plasmids, respectively.
10th: We collected cells 24 hours after transfection, and preserved them in Trizol.
11th: We collected 48 hours after transfection, and then extracted RNA from 2 groups of cells (24 hours and 48 hours).
12th: We did RT-PCR and qPCR with all RNA samples.
13th: We analyzed the data.
 　　Absolute quantification of exosomes (failed) 5th: We cultured 293t cells.
6th: Cells we cultured on 5th July were contaminated by bacteria.
7th: We subcultured another cell line of 293t cells.
9th: We cultured 293t cells in 6 flasks.
10th:We transferred 293T cells with plasmids.
11th: We collected exosomes 24 hours after transfection.
12th: A part of cells died, failed to collect 48-hour exosomes.
13th: We examined protein concentration of 24-hour exosomes, started to extract RNA from 24h-cells and 24h-exosomes and preserved the rough RNA extract solution with isopropyl alcohol at 4℃ overnight.
14th：We continued finish RNA extraction, then did RT-PCR and qPCR.
15th: We analyzed data.
 　　Luciferase Assay 11th: Construction of plasmids: obtain vector and segment.
12th: We combined vector and segment with T4 ligase, then transformed the recombined plasmids into E.coli DH5α competent cells and spread them on solid LB culture plates by streak plate method.
13th: No single bacterial colony was found.
14th: No single bacterial colony was found again, redid previous steps(obtain the target segments, combined vector and segment, and then transferred it into E.coli )
15th: Single bacterial colonies were found. We picked single colonies and transferred them into liquid LB culture medium with ampicillin, and shaken overnight at 37℃.
16th: The transformed E.coli cells failed to reproduce in LB culture medium.
 Others 5th: We thawed 293t cells.
6th: We subcultured HepG2 cells.
7th: E.coli cells containing HBSag overexpressed plasmids were transferred respectively into LB culture medium with ampicillin, and shaken overnight at 37℃.
8th: We extracted HBSag overexpressed plasmids from E.coli cells.
11th: We thawed 293t cells and subcultured HepG2 cells. E.coli cells containing GFP plasmids were transferred into LB culture medium with ampicillin, and shaken overnight at 37℃.
12th: We extracted GFP plasmids from E.coli cells.
14th: We thawed HepG2 cells and 293T cells.
15th: We thawed 293T cells.

September 12, 2013 Elastislide - A Responsive jQuery Carousel Plugin

Elastislide - A Responsive jQuery Carousel Plugin
Slicebox - A fresh 3D image slider with graceful fallback

September 5, 2013 Slicebox - A fresh 3D image slider with graceful fallback

August

2013

August 30, 2013 Automatic Image Montage with jQuery

Automatic Image Montage with jQuery
Image Zoom Tour with jQuery

August 23, 2013 Image Zoom Tour with jQuery

August 16, 2013 Circular Content Carousel with jQuery

Circular Content Carousel with jQuery
Portfolio Image Navigation with jQuery

August 9, 2013 Portfolio Image Navigation with jQuery

August 4, 2013 Expanding Fullscreen Grid Portfolio

Expanding Fullscreen Grid Portfolio

July

2013

Content Rotator with jQuery

July 29, 2013 Content Rotator with jQuery

July 22, 2013 Vertical Sliding Accordion with jQuery

Vertical Sliding Accordion with jQuery
Animated Text and Icon Menu with jQuery

July 12, 2013 Animated Text and Icon Menu with jQuery

July 5, 2013 Fullscreen Slideshow with HTML5 Audio and jQuery

Fullscreen Slideshow with HTML5 Audio and jQuery
Sliding Background Image Menu with jQuery

July 3, 2013 Sliding Background Image Menu with jQuery

June

2013

June 9, 2013 Grid Navigation Effects with jQuery

Grid Navigation Effects with jQuery

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