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OCTOBER
Bio-Lab activity proceeds!
Select the day you are interested in to find out the details!
| DATE | ACTIVITY |
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| October, 1st |
97° Bio-Lab - Trasformed plates were chacked(I70, I71, I72 and I73).
Transformation of I55 into E. coli DH5-alpha. It was plated on LB+Amp 100 agar plate and let grow ON at +37°C.
Miniprep and Nanodrop quantification for: I31, I35, I60 (they were also sent sequencing), INTEIN, pSB1C3-1, pSB1C3-2, pSB1C3-3 and I37 (they were no sent sequencing).
Digest and gel run/cut/extraction/quantification of these samples were performed.
ON ligation of: I74: I31 + I35 and INTEIN_C3: INTEIN + pSB1C3.
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| October, 2nd |
98° Bio-Lab - PCR from colony for I55 (eleven colonies were picked).We decided to make glycerol stock for I55-1. Inoculum into 5 ml LB+Amp of the positive culture that was let grow ON at 37°C, 220 rpm.
Miniprep and quantification for I73-1/2/3/4/5/6. 1,5 hours E-P digest (for screening) and gel run.
Transformation of I74 and INTEIN_C3 (1ul) in E. coli TOP10. They were plated on LB+Amp100 and LB+Cm34 agar plates respectively. They were let grow ON at +37°C.
Inoculum of I55-1 into 5 ml LB+Amp. ON growth at +37°C, 220 rpm.
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| October, 3rd |
99° Bio-Lab - Glycerol stock for:I55-1, IC-1/2/3/4-4C5.
Pick of a single non-red colony of INTEIN_C3 and inoculum into 5 ml LB+Cm34. Sample was let grow ON at +37°C, 220 rpm.
Miniprep and quantification of:I55-1, I55-1bis. I55-1bis was digested E-S for about three hours and than gel run/cut.
Colony PCR for I74-1..6. Gel run for PCR samples and I55 (E-S) and cut for this one.
Gel extraction and quantification of I55.
Ligation of I55 with already available DNA: I75: I55 + I37 and I77: I55 + I57.
Inoculum of I35 (last quantification after gel extraction was too poor; we will digest it again E-P) and I54 into 5 ml LB+Amp: ON growth at +37°C, 220 rpm.
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| OCTOBER: WEEK 1 |
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| October, 4th |
100° Bio-Lab - Miniprep and quantification of: INTEIN_C3, I74-8, I54, I35. Digest and gel run/cut/quantification of these samples.
Ligations of I78=I74+INTEIN_C3.
Transormation (1ul) into E. coli TOP10 of:I75, I77. They were plated on LB+Amp agar plates and let grow ON at +37°C, 220 rpm.
Today we start to transfer all parts that will be sent in <partinfo>pSB1C3</partinfo> plasmid.
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| October, 5th |
101° Bio-Lab - Digestions were performed again for I55, I37, I57, I31 and I54. Digestions were let at +37°C ON.
Transformation (1ul) of I78 into E. coli TOP10. In order to check our E. coli TOP10 competent cells we plated them on LB+Amp and Cm agar plates.
ON ligation of I79: I60 + INTEIN_C3.
We are now moving all our parts to pSB1C3 plasmid.
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| October, 6th |
102° Bio-Lab - E. coli TOP10 plated on LB+Amp agar plate showed red colonies!!!
Transformation of available ligations I75 and I77 (1ul) into E. coli TOP10. They were plated on LB+Amp agar plates and let grow ON at +37°C, 220 rpm.
Gel run and cut for ON digestions and Nanodrop quantification of I54 and I55. Ligation of I76=I55+I54.
PCR from colony screening for I78.
We are now moving all our parts to pSB1C3 plasmid.
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| October, 7th |
103° Bio-Lab - I78 screening gel run. Glycerol stock and miniprep for I78-3.
Colony PCR for screening of I79 (we pick eleven colonies).
Transformation (1ul) of I0_1C3, I3_1C3, I7_1C3, I8_1C3, I9_1C3, I12_1C3 and I76 into E. coli DH5-alpha.
They were plated on LB+Cm34 agar plates (except for I76 that was plated on LB+Amp agar plate) and let grow ON at +37°C.
We are now moving all our parts to pSB1C3 plasmid.
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| October, 8th |
104° Bio-Lab - Glycerol stock, miniprep and Nanodrop quantification of I79-2.This DNA was sent to BMR Genomics for sequencing.
We are now moving all our parts to pSB1C3 plasmid.
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| October, 9th |
105° Bio-Lab -A new part was cloned with these ones, named I80. It is the basic part to build self-inducible promoters.
We are now moving all our parts to pSB1C3 plasmid.
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| October, 10th |
106° Bio-Lab - Inoculum into 4 ml LB+Amp of: I75-1/2/3, I76-1/2, I77-1/2/3, I47, I48, I49. They were let grow at +37°C, 220 rpm.
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| OCTOBER: WEEK 2 |
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| October, 11th |
107° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
Inoculum of MC42-A, MC43-A, MG42-B, MG43-A, MC43_flip-a,b,c, MG43_flip-a,b,c, MG1655.
15° Meeting - Updating about Bio-Lab activity. Decisions about poster and presentation.
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| October, 12th |
108° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
Screening PCR.
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| October, 13th |
109° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
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| October, 14th |
110° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
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| October, 15th |
111° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
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| October, 16th |
112° Bio-Lab - We are now moving all our parts to pSB1C3 plasmid.
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| OCTOBER: WEEK 3 |
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| October, 18th |
113° Bio-Lab - We have finally completed the parts transfer to pSB1C3 plasmid. We sent the parts to MIT.
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| October, 19th |
114° Bio-Lab -
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| October, 20th |
115° Bio-Lab -
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| October, 21st |
116° Bio-Lab -
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| October, 22nd |
117° Bio-Lab -
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| October, 23rd |
118° Bio-Lab -
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| OCTOBER: WEEK 4 |
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| October, 25th |
119° Bio-Lab -
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| October, 26th |
120° Bio-Lab -
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| October, 27th |
121° Bio-Lab - Closing wiki
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