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From 2012.igem.org

Week 10 keep moving on

2012/8/20 Monday, August 20, 2012

1. Zhang continues LIU Xiao's point mutation D0 library experiment. According to the instructions of DNA gel extraction kit, she recover enzyme-digested products product pETDuet-BB, put new D0 into it.

2. We receive part K411003 and plate it on Chloramphenicol plates, and incubate.

2012/8/21 Tuesday, August 21, 2012

1. K411003 (Taipei 2010) were adding a series of Theophylline. This experiment is used both for testing parts and fishing for the condition of our synthetic D0 from Genescript's future GFP experiment.

2. The Synergy hybrid reader data is good. You can see the fluorescence intensity increasing with Theophylline concentration.

2012/8/22 Wednesday, August 22, 2012

1. 00:39 in the morning, YU Jianing sends an e-mail to Yan from her hometown, attaching version 1 ODE modeling and some questions about biology meanings.

2. We start our major experiment: construct FA-2X-MS2 and FB-2X-PP7

2.1 Fragment

1. We PCR FA and FB from pEGFP.

2. We PCR MS2 from FAM( from Dr. Delebecque)

3. We PCR PP7 from FBP(from Dr. Delebecque )

Then use gel extraction kit to get these fragment.

2.2 Overlap PCR

First overlap PCR in our lab. We uses precisely designed primers to make FA and MS2 linked with 2X. So does FB and PP7.

3. Zhang goes on with LIU Xiao's point mutation and use Megaprimer PCR methods.

2012/8/23 Thursday, August 23, 2012

1. Yan PCR riboscaffold (synthesis by Genescript) and make it into Part.

2. Zhang continues mutation D0 and enzyme digest the fragment for PCR test.

3. LIU Huachun uses some parts form registry to construct a new EGFP for positive control.

2012/8/24 Friday, August 24, 2012

1. LIU reproduce pSB1C3

2. Yan continues to make part of riboscaffold.

3. Chen use Taq DNA Polymerase (ordinary) to put A onto FA-2X-MS2 and FB-2X-PP7 and link it onto T vector.

2012/8/25 Saturday, August 25, 2012

1. We extracted and amplified the mutation D0 library and then sent it to a sequencing facility

2. LIU's pCDF-EGFP: transform into DH5α (It works pretty well.)

3. Miniprep: Yan's last day's part riboscaffold

2012/8/26 Sunday, August 26, 2012

1. Miniprep: pCDFDuet-EGFP and pColaDuet

2. Part riboscaffold PCR: a little different from its original PCR from pETDuet Genescript. So we should send sequencing for validation.