IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-4: Difference between revisions
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Qu Mingzhi (talk | contribs) |
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#pSB1A2 vector | #pSB1A2 vector | ||
#marker (DL2000 Plus) | #marker (DL2000 Plus) | ||
[[Image:Peking_2007-8-4_Ori-T_pSB1A2mini-prep_double_digesting_test_result(small).jpg]] | |||
==Double digesting test for F-OriT_pSB1A2 & J23066 == | ==Double digesting test for F-OriT_pSB1A2 & J23066 == |
Revision as of 20:00, 5 August 2007
mini-prep F-OriT_pSB1A2
- using Transgen mini plasmid puriflication kit.
- 30µL after purflication
mini-prep F-OriT_pSB1A2
- using Transgen mini plasmid puriflication kit.
- 30µL after purflication
mini-prep double digesting test result
- Digesting F-OriT_pSB1A2 with EcoR1/PstI.
- Digestion system contains:
1 µl 10*H 0.25 µl EcoRI 0.25 µl PstI 5 µl Plasmid 3.5 µl dH20 -------------------------- 10 µl Total
- 37℃ culutre for 3 hours.
- from left to right:
- mini-prep F-OriT_pSB1A2 -1 @ EcoRI/PstI
- mini-prep F-OriT_pSB1A2 -2 @ EcoRI/PstI
- Ori-T fragment
- pSB1A2 vector
- marker (DL2000 Plus)
Double digesting test for F-OriT_pSB1A2 & J23066
- use Double digesting test for gel extraction.
- F-OriT_pSB1A2 digestion system contains:
4 µl 10*H buffer 1 µl SpeI 1 µl PstI 20 µl Plasmid 14 µl dH20 -------------------------- 40 µl Total
- J23066 digestion system contains:
4 µl 10*M buffer 1 µl XbaI 1 µl PstI 20 µl Plasmid 14 µl dH20 -------------------------- 40 µl Total
electrophoresis result before gel extraction
- from left to right:
- F-OriT_pSB1A2 -1 @ SpeI/PstI
- F-OriT_pSB1A2 -2 @ SpeI/PstI
- J23066 -1 @ XbaI/PstI
- J23066 -2 @ XbaI/PstI
- marker