Oriental fruit flies sent to lab and cultivated by large transparent box to avoid oriental fruit flies flying out.

We used both of centrifuge tube and feeding box to feed oriental fruit flies for comparing and ensuring the status of oriental fruit flies that established the independent space for observing oriental fruit flies.

B. dorsalis started to break through the cocoon. We conducted to put B. dorsalis to centrifuge tubes, avoiding the feeding box cannot load all oriental fruit flies. We poured the special agar to centrifuge tubes, and put centrifuges into entrance to catch oriental fruit flies. Then we sealed the centrifuge tubes by cotton.

First time to Conducting Experiment 1

Changing the centrifuge tubes where oriental fruit flies lived. (Avoid the environment being mess.) /p>

Changing the centrifuge tubes where oriental fruit flies lived.

The oriental fruit flies in centrifuge tubes are dead quickly, so we conducted trouble shooting and sent the mail to professors who research oriental fruit flies.

We took the trap to Beitou orchard to try to catch oriental fruit flies.

Second time to Conducting Experiment 1.

Conducting trouble shooting.

Searching the store selling the silkworm for extracting hemolymph which is used to induce pMcl1.

Visiting two farmers working in the orchard to ask the information about oriental fruit flies in the nature environment, and installing the trap of catching oriental fruit flies.

Sent the mail to researchers to ask for making public the content of the mail.

8/28: RFP_TtrpC digested for ligation with pBARGPE1------ the concentration was not high enough

8/28: RFP and TtrpC did PCR to ligase(failure, we change the volume of the primer and Mg2+)

8/28: EL222 and TtrpC ran ligation PCR ------- NC had band

8/28: EL222 and TtrpC ran ligation PCR ------ with remixed primer

8/28: pVP-EL222 and TtrpC PCR ligation ------- NC had bands

8/28: pVP-EL222 and TtrpC PCR ligation ------- used remixed primer

8/29: pVP-EL222 and TtrpC PCR ligation ------- the concentration was not high enough after gel extraction

8/29: EL222 and TtrpC ran ligation PCR ------ the concentration was not high enough after gel extraction

8/31: EL222 and TtrpC ran ligation PCR

8/31: pBAR_RFP digestion(to transformed into meta)

8/31: RFP_TtrpC digested for ligase with pBARGPE1 (the band existed on wrong site)

8/31: pVP-EL222 and TtrpC PCR ligation

8/31: PMCL1 Taq polymerase PCR amplification

9/1: PMCL1 PCR amplification test run

9/1: PMCL1 KOD polymerase PCR amplification

9/1: PMCL1 polymerase PCR amplification

9/1: RFP_TtrpC digested

9/1: pVP-EL222 and TtrpC PCR ligase

9/1: EL222 and TtrpC ran ligation PCR

9/2: EL222-TtrpC digestion

9/2: pBARGPE1 digested for pBAR_EL222_TtrpC construction

9/2: pBAR _RFP_TtrpC transformed into competent cell

9/2: pBAR_PgpdA digested for ligation with EL222_TtrpC

9/2: PMCL1 KOD polymerase PCR amplification

9/2: PMCL1 KOD polymerase PCR amplification

9/3: PMCL1 KOD polymerase PCR amplification

9/3: pBAR_EL222_TtrpC transformed into competent cell

9/3: pBAR_EL222_TtrpC containing cultures used in 3 in 1

9/3: pBAR_PgpdA digested for ligation with EL222_TtrpC

9/3: pBAR_PgpdA_EL222_TtrpC transformed into competent cell

9/4: pBAR_PgpdA_EL222_TtrpC containing cultures used in 3 in 1

9/4: pBAR_PgpdA_EL222_TtrpC extracted from transformed cultures

9/4: pBAR _RFP_TtrpC did plasmid PCR

9/4: PMCL1 KOD polymerase PCR amplification

9/5: pBAR _RFP_TtrpC transformed into competent cell ------ there was no E.coli on the first plate

9/5: mRFP1 amplified with KOD polymerase PCR

9/5: pBAR_PgpdA_EL222_TtrpC amplified with plasmid PCR

9/5: pBAR_EL222_TtrpC checked with restriction enzyme digestion

9/6: EL222-TtrpC digestion

9/6: pBAR_EL222_TtrpC checked with restriction enzyme digestion

9/6: pBAR_PgpdA_EL222_TtrpC checked with restriction digestion (failure)

9/6: pBAR_PgpdA_EL222_TtrpC checked with restriction digestion (we changed enzymes)

9/6: EL222-TtrpC checked with restriction digestion

9/6: EL222-TtrpC digested with restriction enzyme

9/6: BBa_E0040 streaked onto agar plates

9/6: BBa_K118400 resuspended from 2016 iGEM Kit plate 4

9/6: BBa_K118400 transformed into competent cells

9/6: pBAR _RFP_TtrpC transformed into competent cell

9/6: PMCL1 KOD polymerase PCR amplification

9/6: PMCL1 KOD polymerase PCR amplification

9/7: PMCL1 KOD polymerase PCR amplification

9/7: pBAR _RFP_TtrpC transformed into competent cell

9/7: pBAR _RFP_TtrpC did 2 in 1

9/7: pBAR _RFP_TtrpC did 3 in 1

9/7: BBa_E0040 cultures used in 2 in 1

9/7: pBAR_EL222 PCR

9/7: BBa_K118400 containing culture used in 2 in 1

9/7: pVP-EL222 amplified with PCR (use for ligation with both pBAR_PgpdA and PMCL1)

9/8: pVP-EL222 amplified with PCR (use for ligation with both pBAR_PgpdA and PMCL1)

9/8: BBa_K118400 extraction extracted from transformed cultures

9/8: BBa_K118400 amplified with plasmid PCR

9/8: BBa_E0040 extracted from transformed culture

9/8: pBAR _RFP_TtrpC plasmid PCR

9/8: pBAR _RFP_TtrpC digestion check

9/8: pBAR _RFP_TtrpC digestion check

9/8: pBAR _RFP_TtrpC did plasmid extraction

9/8: PMCL1 KOD polymerase PCR amplification

9/9: PMCL1 KOD polymerase PCR amplification

9/9: pBAR _RFP_TtrpC did digestion check

9/9: BBa_E0040 amplified in plasmid PCR

9/9: mRFP1amplified with KOD polymerase PCR

9/9: BBa_E0040 checked with restriction digestion (successful)

9/9: BBa_K118400 checked with restriction digestion (the gel had faulty matrix)

9/9: BBa_K118400 checked with restriction digestion (confirmed, successful)

9/9: BBa_K1184000 use KOD polymerase to amplify the target part KillerRed (try)

9/9: pBAR_PgpdA_EL222_TtrpC checked with restriction digestion

9/9: pBAR_PgpdA_EL222_TtrpC transformed into competent cell( there was no E.coli on our first plate )

9/9: pBARGPE1 digested for pBAR_mRFP1_TtrpC construction

9/10: pBARGPE1 digested for pBAR_PgpdA_EL222_TtrpC construction

9/10: pBAR_PgpdA_EL222_TtrpC transformed into competent cell

9/10: BBa_K1184000 use KOD polymerase to amplify the target part KillerRed (really amplified)

9/10: RFP and TtrpC use primer to ligase

9/10: RFP_TtrpC PCR to amplify the target sequence

9/10: Digested KillerRed fragment and its backbone pBARGPE1 with BamHI and EcoRI for ligation to construct the vector pBAR_KR

9/10: Digested KillerRed fragment and its backbone pBARGPE1 with BamHI and EcoRI again

9/10: Amplified PMCL1 from genome (KOD polymerase)

9/10: Amplified PMCL1 from genome (KOD polymerase)

9/10: Standard part PgpdA amplification PCR with Taq polymerase

9/11: Standard part PgpdA amplification PCR with KOD polymerase

9/11: pBARGPE1 digested for pBAR_PgpdA_EL222_TtrpC construction

9/11: TtrpC KOD PCR amplification

9/11: TtrpC KOD PCR amplification

9/11: pBAR_PgpdA_EL222_TtrpC containing culture used in 3 in 1

9/11: pBAR_KR transformed into competent cell

9/11: pBAR_KR transformed into competent cell

9/12: pBAR_KR performed plasmid extraction and plasmid PCR (successful)

9/12: pBAR_PgpdA_EL222_TtrpC checked with restriction digestion

9/12: pBAR_PgpdA_EL222_TtrpC amplified with plasmid PCR

9/12: Successfully constructed the plasmid pBAR_PgpdA_EL222_TtrpC

9/12: Standard part PgpdA mutation PCR (mutated PstI)

9/12: Standard part PMCL1 amplification PCR with KOD polymerase

9/13: Standard Part PMCL1 mutation PCR (mutated SpeI)

9/13: Standard part TtrpC amplification PCR with KOD polymerase

9/13: pBAR _RFP_TtrpC transformed into competent cell

9/13: RFP_TtrpC PCR to amplify the target sequence

9/13: pBAR _RFP_TtrpC did 3 in 1

9/13: pBAR _RFP_TtrpC digestion check

9/14: pBAR _RFP_TtrpC plasmid extraction

9/14: RFP_TtrpC digested

9/14: K118400 and TtrpC used PCR to amplify the target sequence

9/14: KR_TtrpC used PCR to amplify the target sequence

9/15: KR_TtrpC digested with restriction enzymes

9/15: Standard part TtrpC mutation PCR (mutated XbaI)

9/15: pBARGPE1 digested for pBAR_KR_TtrpC construction

9/16: pBAR_KR_TtrpC transformed into competent cell

9/16: Transformed pBAR_KR_TtrpC into competent cell

9/16: Digested pBAR_KR for transforming into Metarhizium anisopliae

9/16: Extracted plasmid BBa_K118400

9/16: pBAR_EL222_TtrpC did plasmid PCR

9/16: pBAR_EL222_TtrpC did plasmid PCR (confirmed successfully)

9/16: pBAR _RFP_TtrpC did digestion check

9/17: pBAR_RFP_TtrpC did digestion check

9/17: pBAR_EL222_TtrpC plasmid stored and transformed cultures cryopreserved

9/17: pBAR_KR_TtrpC did 3 in 1

9/17: Extracted plasmid pBAR_KR_TtrpC

9/18: Transformed pBAR_RFP_TtrpC into competent cell

9/18: pBAR_RFP_TtrpC did 3 in 1

9/18: pBAR_KR_TtrpC did plasmid PCR and digestion check

9/18: BBa_E0040 did plasmid PCR

9/19: Amplified GFP from plasmid BBa_E0040

9/19: Extracted plasmid BBa_E0040

9/20: KR_TtrpC digestion

9/20: Transformed pBAR_KR_TtrpC into competent cell

9/20: pBAR_KR_TtrpC did 3 in 1

9/20: Extracted plasmid pBAR_KR_TtrpC

9/20: pBAR_KR_TtrpC did digestion check

9/20: Extracted plasmid pBAR_RFP_TtrpC

9/20: pBAR_RFP_TtrpC did digestion check

9/20: RFP_TtrpC digestion

9/21: pBAR_KR_TtrpC did digestion check

9/21: pBAR_RFP_TtrpC did digestion check

9/21: pBAR_RFP_TtrpC did 2 in 1

9/21: pBARGPE1 and RFP_TtrpC digestion

9/22: Amplified RFP_TtrpC with KOD polymerase

9/22: Extracted plasmid pBAR_KR_TtrpC

9/22: pBAR_KR_TtrpC did plasmid PCR

9/22: pMCL1 and pBAR_KR_TtrpC digestion (pMCL1 was not digested successfully)

9/22: pBAR_PgpdA_EL222_TtrpC digested for transforming into Metarhizium anisopliae

9/23: Amplified pMCL1 with KOD polymerase

9/24: pBAR_RFP_TtrpC did digestion check

9/24: Extracted plasmid pBAR_RFP_TtrpC (from 9/5 plate)

9/25: Transformed pBAR_RFP_TtrpC into competent cell

9/25: RFP_TtrpC digestion

9/25: Amplified RFP-TtrpC with KOD polymerase

9/25: Amplified RFP_TtrpC with KOD polymerase

9/25: pMCL1 and pBAR_KR_TtrpC digestion

9/25: Transformed pBAR_pMCL1_KR_TtrpC into competent cell

9/25: Transformed pBAR_pMCL1_KR_TtrpC into competent cell

9/25: pBAR_pMCL1_KR_TtrpC did 3 in 1

9/26: Extracted plasmid pBAR_pMCL1_KR_TtrpC

9/26: pBAR_RFP_TtrpC did 2 in 1

9/26: Extracted plasmid pBAR_RFP_TtrpC

9/26: pBAR_RFP_TtrpC did digestion check

9/26: pBAR_RFP_TtrpC did digestion check

9/26: Amplified pMCL1 with KOD polymerase

9/27: Amplified pMCL1 with KOD polymerase

9/28: pBAR_pMCL1_KR_TtrpC did digestion check

9/28: pBAR_RFP_TtrpC did plasmid PCR

9/28: Successfully constructed the plasmid pBAR_RFP_TtrpC

9/29: Amplified pMCL1 with KOD polymerase

9/30: pBAR_pMCL1_KR_TtrpC did digestion check

9/30: Received the junction primer of GFP-TtrpC

9/30: GFP_TtrpC ran ligation PCR

10/1: GFP_TtrpC ran ligation PCR

10/2: GFP_TtrpC digested for ligation with pBARGPE1

10/2: Transformed pBAR_GFP_TtrpC into competent cells

10/2: pBARGPE1 digested for pBAR_GFP_TtrpC construction

10/2: pBARGPE1 resuspended (from cryopreservation)

10/3: Extracted plasmid pBARGPE1

10/3: pBAR_GFP_TtrpC containing cultures used in 2 in 1

10/3: Extracted plasmid pBAR_GFP_TtrpC

10/3: pBAR_GFP_TtrpC digestion check(failure)

10/3: pBAR_GFP_TtrpC PCR check(failure)

10/3: Transformed pBAR_GFP_TtrpC into competent cells

10/3: pBARGPE1 checked with restriction digestion (correct)

10/4: pBARGPE1 plasmid stored

10/4: pBAR_GFP_TtrpC containing cultures used in 2 in 1

10/4: pBAR_GFP_TtrpC plasmid extraction

10/4: pBAR_GFP_TtrpC digestion check