Team:NUDT CHINA/Notebook

NOTEBOOK

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CULTURE&SQUENCE: Related parts in Kit

After long time waiting and discussing, we can start our experience now.

2015.5.16
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ACTIVITY: Move the lab.

Because of various restrictions, we move our lat to an idle lab.

2015.5.31

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PCR: IaaM&IaaH

We got fragments of IAAM & IAAH to do their standardization next.

2015.6.7

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STANDARDIZAE: IaaM&IaaH

At this point, we could use them convenient.

2015.6.14

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STANDARDIZAE: TALE1

This fragment came from NJU_CHINA, we standardized it ourselves.

2015.6.21
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STANDARDIZAE: SCAF1, SCAF2, SCAF3, TALE2, TALE3

We got SCAF1, SCAF2, SCAF3 a few days ago, and followed the NJU_CHINA produced TALE2,TALE3. Nowadays we standardize them.

2015.6.28
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STANDARDIZAE: Plac+RBS30

Plac+RBS30 is an useful fragment, we standardized it in order to use it.

2015.6.29

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Device(No.1): GFP–N1

We finished the first device. This device is one of negative control for GFP.

2015.7.25
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Device(No.2): GFP-S1

We finished the second device which is the experimental group.

2015.7.27
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Device(No.3): GFP-N2

This device is also one of negative control for GFP.

2015.7.31

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STANDARDIZAE: RBS30

The RBS30 in kit is mistake. We rework it by annealing.

2015.8.12
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BUILD: IAA(+/+)N
2015.8.17
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BUILD: IAA(+/+)S2/S3, GFP P
2015.8.19
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BUILD: GFP-S2, GFP-S3, GFP-P
2015.8.20

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REBUILD: GFP-S1, GFP-S2, GFP-S3, GFP-N1, GFP-N2, IAA, IAA-S2, IAA-S3.

From the result of DNA sequencing, we found the GFP2 from kits had 1 bp too many. Because of this, GFP2 expression is abnormal. We rebuild the GFP2 by PCR, and rebuild the devices. Also from the result of DNA sequencing, the RBS30 in IAA, IAA-S2 and IAA-S3 were disappeared. We rebuild them too.

2015.8.26 — 2015.9.6
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ChIP: GFP series

This experiment proved the function of TALE.

2015.9.4 - 2015.9.6
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ELISA: IAA, IAA-S2, IAA-S3

We found that the production for auxin increased when there have SCAF. And the close two enzymes got, the higher efficiency can be.

2015.9.8
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RT: IAA, IAA-S2, IAA-S3

We could see that the production for RNA got close.

2015.9.9