From 2010.igem.org
iGEM Panama
October 18
Inoculation
We inoculated 5 colonies in liquid LB + amp medium
-3 ml medium + 1 colony
We incubated them overnight at 37C with shaking
Prepare other aliquot of primer for the amplification of the gene Rh1ab
Label the tubes well
Homemade Miniprep
5 samples
"Dirty" Mini Preps
p1: 50mM Tris-HCl, pH8.0; P2: 200mM NaOH, 1% SDS; P3: 3.0M KAc, pH5.5
1. Spin down 1.5 ml cells at 14000 rpm for 2 minutes
Discard supernatant.
2. Resuspend in 300 ul Buffer P1 (with RNAse A)
3. Add 300 ul P2. Mix by inverting. Incubate at room temperature for 5 minutes
4. Add 300 ul of cold Buffer P3.
Mix by inverting, incubate on ice for 5 minutes
5. Spin 14000rpm, 10 minutes
6. Transfer supernatant to a new tube.
Extract with 500 ul Phenol:Chloroform:Isoamyl.
Spin 14000rpm for 5 minutes
7. Transfer 800 ul of the aqueous phase to a new tube
8. Add 0.7 volumes of isopropanol (560 ul)
Mix by inverting
Spin 14000rpm for 5 minutes at 4C
9. Remove isopropanol. Wash with EtOH 70%. Spin for 2-5 minutes at 14000rpm at 4C.
Remove all EtOH.
10. Air dry. Resuspend in 20 ul TE Buffer.
Electrophoresis: Miniprep of plasmid +Rh1ab on a 1% agarose with 1x TBE
Concentration of sample 5, according to Qubit fluorometer = 128ng/ul
Possible cause of the bands being so faint: We didn't let the pellet resuspend for an hour in step 10
May
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_May_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_May_2010&action=edit 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_May_2010&action=edit 3 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/4_May_2010&action=edit 4 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/5_May_2010&action=edit 5 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_May_2010&action=edit 6 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/7_May_2010&action=edit 7 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/8_May_2010&action=edit 8 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/9_May_2010&action=edit 9 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_May_2010&action=edit 10 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/11_May_2010&action=edit 11 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_May_2010&action=edit 12 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_May_2010&action=edit 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_May_2010&action=edit 14 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/15_May_2010&action=edit 15 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_May_2010&action=edit 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_May_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_May_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_May_2010&action=edit 19 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_May_2010&action=edit 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_May_2010&action=edit 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_May_2010&action=edit 22 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/23_May_2010&action=edit 23 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/24_May_2010&action=edit 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_May_2010&action=edit 25 ]
[http://2010.igem.org/Team:Panama/26_May_2010 26 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_May_2010&action=edit 27 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_May_2010&action=edit 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_May_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_May_2010&action=edit 30 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_May_2010&action=edit 31 ]
June
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_June_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_June_2010&action=edit 2 ]
[http://2010.igem.org/Team:Panama/3_June_2010 3 ]
[http://2010.igem.org/Team:Panama/4_June_2010 4 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/5_June_2010&action=edit 5 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_June_2010&action=edit 6 ]
[http://2010.igem.org/Team:Panama/7_June_2010 7 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/8_June_2010&action=edit 8 ]
[http://2010.igem.org/Team:Panama/9_June_2010 9 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_June_2010&action=edit 10 ]
[http://2010.igem.org/Team:Panama/11_June_2010 11 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_June_2010&action=edit 12 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_June_2010&action=edit 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_June_2010&action=edit 14 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/15_June_2010&action=edit 15 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_June_2010&action=edit 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_June_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_June_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_June_2010&action=edit 19 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_June_2010&action=edit 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_June_2010&action=edit 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_June_2010&action=edit 22 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/23_June_2010&action=edit 23 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/24_June_2010&action=edit 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_June_2010&action=edit 25 ]
[http://2010.igem.org/Team:Panama/26_June_2010 26 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_June_2010&action=edit 27 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_June_2010&action=edit 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_June_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_June_2010&action=edit 30 ]
July
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_July_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_July_2010&action=edit 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_July_2010&action=edit 3 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/4_July_2010&action=edit 4 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/5_July_2010&action=edit 5 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_July_2010&action=edit 6 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/7_July_2010&action=edit 7 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/8_July_2010&action=edit 8 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/9_July_2010&action=edit 9 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_July_2010&action=edit 10 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/11_July_2010&action=edit 11 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_July_2010&action=edit 12 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_July_2010&action=edit 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_July_2010&action=edit 14 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/15_July_2010&action=edit 15 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_July_2010&action=edit 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_July_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_July_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_July_2010&action=edit 19 ]
[http://2010.igem.org/Team:Panama/20_July_2010 20 ]
[http://2010.igem.org/Team:Panama/21_July_2010 21 ]
[http://2010.igem.org/Team:Panama/22_July_2010 22 ]
[http://2010.igem.org/Team:Panama/23_July_2010 23 ]
[http://2010.igem.org/Team:Panama/24_July_2010 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_July_2010&action=edit 25 ]
[http://2010.igem.org/Team:Panama/26_July_2010 26 ]
[http://2010.igem.org/Team:Panama/27_July_2010 27 ]
[http://2010.igem.org/Team:Panama/28_July_2010 28 ]
[http://2010.igem.org/Team:Panama/29_July_2010 29 ]
[http://2010.igem.org/Team:Panama/30_July_2010 30 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_July_2010&action=edit 31 ]
August
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_August_2010&action=edit 1 ]
[http://2010.igem.org/Team:Panama/2_August_2010 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_August_2010&action=edit 3 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/4_August_2010&action=edit 4 ]
[http://2010.igem.org/Team:Panama/5_August_2010 5 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_August_2010&action=edit 6 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/7_August_2010&action=edit 7 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/8_August_2010&action=edit 8 ]
[http://2010.igem.org/Team:Panama/9_August_2010 9 ]
[http://2010.igem.org/Team:Panama/10_August_2010 10 ]
[http://2010.igem.org/Team:Panama/11_August_2010 11 ]
[http://2010.igem.org/Team:Panama/12_August_2010 12 ]
[http://2010.igem.org/Team:Panama/13_August_2010 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_August_2010&action=edit 14 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/15_August_2010&action=edit 15 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_August_2010&action=edit 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_August_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_August_2010&action=edit 18 ]
[http://2010.igem.org/Team:Panama/19_August_2010 19 ]
[http://2010.igem.org/Team:Panama/20_August_2010 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_August_2010&action=edit 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_August_2010&action=edit 22 ]
[http://2010.igem.org/Team:Panama/23_August_2010 23 ]
[http://2010.igem.org/Team:Panama/24_August_2010 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_August_2010&action=edit 25 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/26_August_2010&action=edit 26 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_August_2010&action=edit 27 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_August_2010&action=edit 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_August_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_August_2010&action=edit 30 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_August_2010&action=edit 31 ]
September
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_September_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_September_2010&action=edit 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_September_2010&action=edit 3 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/4_September_2010&action=edit 4 ]
[http://2010.igem.org/Team:Panama/5_September_2010 5 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_September_2010&action=edit 6 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/7_September_2010&action=edit 7 ]
[http://2010.igem.org/Team:Panama/8_September_2010 8 ]
[http://2010.igem.org/Team:Panama/9_September_2010 9 ]
[http://2010.igem.org/Team:Panama/10_September_2010 10 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/11_September_2010&action=edit 11 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_September_2010&action=edit 12 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_September_2010&action=edit 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_September_2010&action=edit 14 ]
[http://2010.igem.org/Team:Panama/15_September_2010 15 ]
[http://2010.igem.org/Team:Panama/16_September_2010 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_September_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_September_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_September_2010&action=edit 19 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_September_2010&action=edit 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_September_2010&action=edit 21 ]
[http://2010.igem.org/Team:Panama/22_September_2010 22 ]
[http://2010.igem.org/Team:Panama/23_September_2010 23 ]
[http://2010.igem.org/Team:Panama/24_September_2010 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_September_2010&action=edit 25 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/26_September_2010&action=edit 26 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_September_2010&action=edit 27 ]
[http://2010.igem.org/Team:Panama/28_September_2010 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_September_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_September_2010&action=edit 30 ]
October
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_October_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_October_2010&action=edit 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_October_2010&action=edit 3 ]
[http://2010.igem.org/Team:Panama/4_October_2010 4 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/5_October_2010&action=edit 5 ]
[http://2010.igem.org/Team:Panama/6_October_2010 6 ]
[http://2010.igem.org/Team:Panama/7_October_2010 7 ]
[http://2010.igem.org/Team:Panama/8_October_2010 8 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/9_October_2010&action=edit 9 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_October_2010&action=edit 10 ]
[http://2010.igem.org/Team:Panama/11_October_2010 11 ]
[http://2010.igem.org/Team:Panama/12_October_2010 12 ]
[http://2010.igem.org/Team:Panama/13_October_2010 13 ]
[http://2010.igem.org/Team:Panama/14_October_2010 14 ]
[http://2010.igem.org/Team:Panama/15_October_2010 15 ]
[http://2010.igem.org/Team:Panama/16_October_2010 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_October_2010&action=edit 17 ]
[http://2010.igem.org/Team:Panama/18_October_2010 18 ]
[http://2010.igem.org/Team:Panama/19_October_2010 19 ]
[http://2010.igem.org/Team:Panama/20_October_2010 20 ]
[http://2010.igem.org/Team:Panama/21_October_2010 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_October_2010&action=edit 22 ]
[http://2010.igem.org/Team:Panama/23_October_2010 23 ]
[http://2010.igem.org/Team:Panama/24_October_2010 24 ]
[http://2010.igem.org/Team:Panama/25_October_2010 25 ]
[http://2010.igem.org/Team:Panama/26_October_2010 26 ]
[http://2010.igem.org/Team:Panama/27_October_2010 27 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_October_2010&action=edit 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_October_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_October_2010&action=edit 30 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_October_2010&action=edit 31 ]