Team:UNIPV-Pavia/Calendar/October

From 2010.igem.org

(Difference between revisions)

Revision as of 08:55, 8 October 2010

OCTOBER

Bio-Lab activity proceeds!

Select the day you are interested in to find oout the details!

DATE	ACTIVITY

October, 1st	101° 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.
October, 2nd	102° 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.
October, 3rd	103° 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.
OCTOBER: WEEK 1
October, 4th	104° 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.
October, 5th	105° 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.
October, 6th	106° 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.
October, 7th	107° 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.
October, 8th	108° Bio-Lab -
October, 9th	109° Bio-Lab -
OCTOBER: WEEK 2
October, 11th	110° Bio-Lab -
October, 12th	111° Bio-Lab -
October, 13th	112° Bio-Lab -
October, 14th	113° Bio-Lab -
October, 15th	114° Bio-Lab -
October, 16th	115° Bio-Lab -
OCTOBER: WEEK 3
October, 18th	116° Bio-Lab -
October, 19th	117° Bio-Lab -
October, 20th	118° Bio-Lab -
October, 21st	119° Bio-Lab -
October, 22nd	120° Bio-Lab -
October, 23rd	121° Bio-Lab -
OCTOBER: WEEK 4
October, 25th	122° Bio-Lab -
October, 26th	123° Bio-Lab -
October, 27th	124° Bio-Lab - Closing wiki

@@ Line 72: / Line 72: @@
 <tr><th align="center" width="12%" colspan="2"><b>OCTOBER: WEEK 1</b></th></tr>
 <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/October/settimana1 #October, 4th|October, 4th]]</td><td align="left">
-° Bio-Lab -
+° 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.
 </td></tr>
 <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/October/settimana1 #October, 5th|October, 5th]]</td><td align="left">
-° Bio-Lab -
+° 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.
 </td></tr>
 <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/October/settimana1 #October, 6th|October, 6th]]</td><td align="left">
-° Bio-Lab -
+° 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.
 </td></tr>
 <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/October/settimana1 #October, 7th|October, 7th]]</td><td align="left">
-° Bio-Lab -
+° 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.
 </td></tr>
 <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/October/settimana1 #October, 8th|October, 8th]]</td><td align="left">