IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-12: Difference between revisions
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Qu Mingzhi (talk | contribs) (New page: =Tandem Ori-T by Qu Mingzhi & Ren Ze= ==colony PCR Test for R751, pSC101(II)== *Test Lb- R751 plate, Lb- pSC101 have the correct plasmid. *Test plate:LB- R751, Lb- pSB101, Tc+ R751-pSC101...) |
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*Test Lb- R751 plate, Lb- pSC101 have the correct plasmid. | *Test Lb- R751 plate, Lb- pSC101 have the correct plasmid. | ||
*Test plate:LB- R751, Lb- pSB101, | *Test plate:LB- R751, Lb- pSB101, R751-pSC101 X Dh5α-R0040, R751 X Dh5α-R0040 | ||
*primer :R751 OriT primer, pSC101 primer. | *primer :R751 OriT primer, pSC101 primer. | ||
*according to <Colony PCR STANDARD PROTOCOL> | *according to <Colony PCR STANDARD PROTOCOL> | ||
Revision as of 00:34, 20 August 2007
Tandem Ori-T by Qu Mingzhi & Ren Ze
colony PCR Test for R751, pSC101(II)
- Test Lb- R751 plate, Lb- pSC101 have the correct plasmid.
- Test plate:LB- R751, Lb- pSB101, R751-pSC101 X Dh5α-R0040, R751 X Dh5α-R0040
- primer :R751 OriT primer, pSC101 primer.
- according to <Colony PCR STANDARD PROTOCOL>
- PCR system contains(each well):
0.5 µl Primer 1(100uM) 0.5 µl Primer 2 2 µl dNTP(2.5uM) 2.5 µl 10X Taq Buffer 0.25 µl Taq 19 µl dH20 1 µl template -------------------------- ~25 µl Total
- PCR program condition 1: 94℃ 5min, 94℃ 30s, 53℃ 30s, 72℃ 45s, Go to step 2 for 29 times, 72℃ 10min, 4℃ end.
- PCR program condition 2: 94℃ 5min, 94℃ 30s, 57℃ 30s, 72℃ 45s, Go to step 2 for 29 times, 72℃ 10min, 4℃ end .
electrophorsis result
- top:from left to right
- 1-4 pSC101
- 5-8 R751
- 9-16 R751-pSC101
- 17 Maker(DL2000 plus)
- botton:from left to right
- 18-21 Dh5α-pSC101
- 22-23 Dh5α-R0040
