Team:Wageningen UR/Journal/week11

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(Difference between revisions)
(→week 11: 9 july - 15 july)
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* Production of a HepB PCR fragment with pre- and suffix by means of a PCR using primers annealing to a plasmid containing the Hepatitis B core protein encoding gene. These primers also have an overhang that encode for pre- and suffix
* Production of a HepB PCR fragment with pre- and suffix by means of a PCR using primers annealing to a plasmid containing the Hepatitis B core protein encoding gene. These primers also have an overhang that encode for pre- and suffix
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'''DLS boundary experiment'''
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 +
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''11th - 12th July''
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<ul>
 +
<li>Ultracentrifuge 2 times</li>
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<li>Followed protocol of purification CCMV</li>
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<li>Total end volume is 6 ml</li>
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</ul>
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 +
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''13th July''
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<ul>
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<li>FPLC with the S400 column</li>
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<li>1 run with 4 ml sample</li>
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<li>A very good separation also high concentration</li>
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</ul>
 +
 +
 +
''14th July''
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<ul>
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<li>DLS Measurement: 20 runs, 45 seconds</li>
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</ul>
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 +
----
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[[https://2012.igem.org/Team:Wageningen_UR/Journal/week10 previous week]]          [[https://2012.igem.org/Team:Wageningen_UR/Journal/week12 next week]]
[[https://2012.igem.org/Team:Wageningen_UR/Journal/week10 previous week]]          [[https://2012.igem.org/Team:Wageningen_UR/Journal/week12 next week]]

Revision as of 09:14, 24 September 2012

week 11: 9 july - 15 july

Office work

Lab work

TuYV

  • Two (non-readthrough) gene variants 1(TuYV coat protein) and 1H(TuYV coat protein with his-tag) were digested and ligated into pSB1A3 linearized backbone.


Written by: Han Yue

PLRV

The start of the lab work on PLRV

We went out to the potato fields near Wageningen to look for PLRV infected potatoes. We thought we found some infected plants, based on the primary symptoms namely smaller sizes of the plants and curled leafs (figure 1).

Figure 1:...

We contacted Kees Bus and Chris Cuperus, who are experts at the Wageningen University on potato diseases. Judging the pictures, they concluded that these plants were probably not infected by PLRV.

This week we started trial experiments for RNA isolation from potato plants. We used a RNA isolation protocols using Trizol (see protocol page). The RNA product concentration was very high (upto 3500 ng/µL).

Figure 2:...

RNA isolate purity checked. RNA is of very good quality. Note that smaller rRNA bands visible in the leaf sample are derived from plant organelles such as plastids and chloroplasts

HepB

  • Production of a HepB PCR fragment with pre- and suffix by means of a PCR using primers annealing to a plasmid containing the Hepatitis B core protein encoding gene. These primers also have an overhang that encode for pre- and suffix

DLS boundary experiment


11th - 12th July

  • Ultracentrifuge 2 times
  • Followed protocol of purification CCMV
  • Total end volume is 6 ml


13th July

  • FPLC with the S400 column
  • 1 run with 4 ml sample
  • A very good separation also high concentration


14th July

  • DLS Measurement: 20 runs, 45 seconds

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