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27645 GACGACGCCTCTGC(CA/GG)CCG(C/T)TTCCAAG(A/G)C(G/C)GAG(A/G)CC(AG/GC)
27646 GACGACGCCTCTGCCAC(c/g)g(ct/tc)tccaagacggagacc(ag/gc)
27644 GACGACGCC(T/G)C(T/G)GCC(a/g)ccg(c/t)ttccaag(a/g)cggag(a/g)cc
27649 and 27648 GACGACGCCTC(t/g)gcc(a/g)ccg(c/t)ttccaag(a/g)cgga(a/g)
27647 GACGACGC(ct/gg)c(t/c)gccaccg(c/t)t(t/g)ccaagacggagacc(ag/gc)
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
24408 | GCT TAG TGA GGG GAC ATG CT | Forward | A | |||
24412 | ACT GAG GGA GCC TGC ATG A | Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
24408 | 0.50 uM |
24412 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |
Stock Number | Strain Name |
---|---|
027644 | STOCK Rhotm1Jlem Tg(Rho*)1PRieke/J |
027645 | STOCK Rhotm1Jlem Tg(Rho*)2PRieke/J |
027646 | STOCK Rhotm1Jlem Tg(Rho*)5PRieke/J |
027647 | STOCK Rhotm1Jlem Tg(Rho*)STM2Rieke/J |
027648 | STOCK Rhotm1Jlem Tg(Rho*)TTM1Rieke/J |
027649 | STOCK Rhotm1Jlem Tg(Rho*)TTM4Rieke/J |
6 strains use this protocol |