"

 

From 2008.igem.org

>送受信班

>Protocols

Contents 1 Week 1 1.1 17 August, 2008 1.2 18 August, 2008 1.3 19 August, 2008 1.4 20 August, 2008 1.5 21 August, 2008 1.6 22 August, 2008 1.7 23 August, 2008 1.8 >next week

Week 1

17 August, 2008

Transformation

competent cells : XL10G

• BBa_R0040(2008) [1] -> No colonies on plates
• BBa_C0070(2008) [2] -> No colonies on plates
• BBa_C0076(2008) [3] -> No colonies on plates
• BBa_C0077(2008) [4] -> No colonies on plates
• BBa_C0078(2008) [5] -> No colonies on plates
• BBa_C0079(2008) [6] -> No colonies on plates

18 August, 2008

PCR

• BBa_C0070(2008) [7]
• BBa_C0076(2008) [8]
• BBa_C0077(2008) [9]
• BBa_C0078(2008) [10]
• BBa_C0079(2008) [11]
• BBa_F2620(2007) [12]･･･control

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase	1
Thermopol Buffer	10
dH2O	68
TOTAL	100μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0070(2008) [13] ->None BBa_C0076(2008) [14] ->None BBa_C0077(2008) [15] ->None BBa_C0078(2008) [16] ->None BBa_C0079(2008) [17] ->None BBa_F2620(2007) [18]･･･control ->OK

Transformation

competent cells : XL10G

• BBa_C0070(2007) [19]
• BBa_C0070(2006) [20]
• BBa_C0076(2007) [21]
• BBa_C0076(2006) [22] -> No colonies on plates(19/8)
• BBa_C0078(2007) [23]
• BBa_C0078(2006) [24]
• BBa_C0170(2007) [25]
• BBa_C0170(2006) [26]
• BBa_C0178(2007) [27]
• BBa_C0178(2006) [28]

--->(20/8)Mini prep

19 August, 2008

PCR

• BBa_I9026(2007) [29]
• BBa_I9026(2006) [30]
• BBa_I9030(2007) [31]
• BBa_I9030(2006) [32]
• BBa_S3154(2007) [33]
• BBa_S3154(2006) [34]
• BBa_F2620(2007) [35]･･･control

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase VENT	1
Thermopol Buffer	5
dH2O	28
TOTAL	50μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_I9026(2007) [36] ->None BBa_I9026(2006) [37] ->None BBa_I9030(2007) [38] ->None BBa_I9030(2006) [39] ->None BBa_S3154(2007) [40] ->None BBa_S3154(2006) [41] ->None BBa_F2620(2007) [42]･･･control ->OK

20 August, 2008

(18/8)--->Mini prep

• BBa_C0070(2007) [43]
• BBa_C0070(2006) [44]
• BBa_C0076(2007) [45]
• BBa_C0078(2007) [46]
• BBa_C0078(2006) [47]
• BBa_C0170(2007) [48]
• BBa_C0170(2006) [49]
• BBa_C0178(2007) [50]
• BBa_C0178(2006) [51]

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0076(2007) [52] -> None BBa_C0078(2007) [53] -> None BBa_C0078(2006) [54] -> None

Sample DNA 10 Loading Dye 4 dH2O 10 TOTAL 24μL From left; BBa_C0078(2007) [55] -> None BBa_C0078(2006) [56] -> OK BBa_C0076(2007) [57] -> None BBa_C0070(2007) [58] -> OK BBa_C0070(2006) [59] -> OK

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0170(2007) [60] -> OK BBa_C0170(2006) [61] -> OK BBa_C0178(2007) [62] -> OK BBa_C0178(2006) [63] -> OK

--->Digestion test

• BBa_C0170(2007) [64]
• BBa_C0170(2006) [65]
• BBa_C0178(2007) [66]
• BBa_C0178(2006) [67]

Sample No	1	2	3	4	5	6	7	8
BBa_C0170(2006) [68]	3	-	-	-	3	-	-	-
BBa_C0170(2007) [69]	-	3	-	-	-	3	-	-
BBa_C0178(2006) [70]	-	-	3	-	-	-	3	-
BBa_C0178(2007) [71]	-	-	-	3	-	-	-	3
EcoRⅠ	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1
PstⅠ	-	-	-	-	0.1	0.1	0.1	0.1
Buffer EcoRⅠ	1	1	1	1	1	1	1	1
BSA	1	1	1	1	1	1	1	1
dH2O	4.9	4.9	4.9	4.9	4.8	4.8	4.8	4.8
TOTAL	10	10	10	10	10	10	10	10

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; (Single Digestion 1~4) BBa_C0170(2006) [72] -> OK BBa_C0170(2007) [73] -> OK BBa_C0178(2006) [74] -> OK BBa_C0178(2007) [75] -> OK (Double Digestion 1~4) BBa_C0170(2006) [76] -> OK BBa_C0170(2007) [77] -> OK BBa_C0178(2006) [78] -> OK BBa_C0178(2007) [79] -> OK

21 August, 2008

(20/8)--->PCR

• BBa_C0070(2007) [80]
• BBa_C0070(2006) [81]
• BBa_C0076(2007) [82]
• BBa_C0078(2007) [83]
• BBa_C0078(2006) [84]

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase TAQ	1
Thermopol Buffer	5
dH2O	28
TOTAL	50μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0078(2007) [85] -> None BBa_C0078(2006) [86] -> None BBa_C0076(2007) [87] -> None BBa_C0070(2007) [88] -> OK BBa_C0070(2006) [89] -> OK

Transformation

competent cells : XL10G

• BBa_S03154(2007) [90]
• BBa_S03154(2006) [91]
• BBa_I9026(2007) [92]
• BBa_I9026(2006) [93]
• BBa_I9030(2007) [94]
• BBa_I9030(2006) [95]

--->(23/8)Digestion

22 August, 2008

Transformation

competent cells : XL10G

• BBa_J13002(2007) [96]
• BBa_J13002(2006) [97]
• BBa_J04500(2007) [98]
• BBa_J04500(2006) [99]

(Locate & 2 Punch ~iyama ver.~)

• BBa_C0076(2008) [100] -> No colonies on plates
• BBa_C0077(2008) [101] -> No colonies on plates
• BBa_C0078(2008) [102] -> No colonies on plates
• BBa_J04500(2008) [103] -> No colonies on plates

--->(24/8)Mini prep

23 August, 2008

(21/8)--->Mini prep

• BBa_S03154(2007) [104]
• BBa_S03154(2006) [105]
• BBa_I9026(2007) [106]
• BBa_I9026(2006) [107]
• BBa_I9030(2007) [108]
• BBa_I9030(2006) [109]

--->Digestion test

• BBa_S03154(2007) [110]
• BBa_S03154(2006) [111]
• BBa_I9026(2007) [112]
• BBa_I9026(2006) [113]
• BBa_I9030(2007) [114]
• BBa_I9030(2006) [115]
• BBa_J04500(2007) [116]

Sample No	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
BBa_I9030(2007) [117]	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-
BBa_I9030(2006) [118]	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-
BBa_S03154(2007) [119]	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-
BBa_S03154(2006) [120]	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-
BBa_I9026(2007) [121]	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-
BBa_I9026(2007) [122]	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-
BBa_J04500(2007) [123]	-	-	-	-	-	-	1	-	-	-	-	-	-	3	-	-	-	-	-	-	3
EcoRⅠ	-	-	-	-	-	-	-	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1
PstⅠ	-	-	-	-	-	-	-	-	-	-	-	-	-	-	0.1	0.1	0.1	0.1	0.1	0.1	0.1
Buffer EcoRⅠ	-	-	-	-	-	-	-	0.9	0.9	0.9	0.9	0.9	0.9	0.9	0.8	0.8	0.8	0.8	0.8	0.8	0.8
BSA	-	-	-	-	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	1	1
dH2O	-	-	-	-	-	-	-	5	5	5	5	5	5	5	5	5	5	5	5	5	5
TOTAL	3	3	3	3	3	3	1	10	10	10	10	10	10	10	10	10	10	10	10	10	10

--->Gel Check

Sample No 1~6 7 Sample DNA 3 1 Loading Dye 1 1 dH2O 2 4 TOTAL 6 6 From left; Sanple No. 1~7 BBa_S03154(2007) [124] BBa_S03154(2006) [125] BBa_I9026(2007) [126] BBa_I9026(2006) [127] BBa_I9030(2007) [128] BBa_I9030(2006) [129] BBa_J04500(2007) [130]

Sample No 8~21 Sample DNA 10 Loading Dye 2 TOTAL 12 From left; (Single Digestion) Sanple No. 8~14 BBa_S03154(2007) [131] -> OK BBa_S03154(2006) [132] -> OK BBa_I9026(2007) [133] -> OK BBa_I9026(2006) [134] -> OK BBa_I9030(2007) [135] -> OK BBa_I9030(2006) [136] -> OK BBa_J04500(2007) [137] -> OK (Double Digestion) Sample No. 15~21 BBa_S03154(2007) [138] -> OK BBa_S03154(2006) [139] -> OK BBa_I9026(2007) [140] -> OK BBa_I9026(2006) [141] -> OK BBa_I9030(2007) [142] -> OK BBa_I9030(2006) [143] -> OK BBa_J04500(2007) [144] -> OK?

Digestion test

BBa_J04500(2007:old-sample)[145]

Sample No	1	2	3
Sample DNA	3	3	3
EcoRⅠ	-	0.1	0.1
SpeⅠ	-	-	0.1
EcoRⅠ Buffer	-	0.9	0.8
BSA	-	1	1
dH2O	-	5	5
TOTAL	3	10	10

--->Gel Check

Sample No 1 2 3 Sample DNA 3 10 10 Loading Dye 1 2 2 dH2O 2 - - TOTAL 6 12 12 From left; BBa_J04500(2007) [146] -> OK BBa_J04500(2007) [147]Single Digestion -> OK BBa_J04500(2007) [148]Double Digestion -> Bad

>next week

ホーム	メンバー紹介	プロジェクト紹介	Parts Submitted to the Registry	モデリング	ノート

Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B2/week_1"

• Recent changes
• What links here
• Related changes
• Special pages
• My preferences

• Printable version
• Permanent link
• Privacy policy
• Disclaimers