Table 3
Addition of the C-tail’s reaction
Reagent
One reaction
Sheared DNA
500 ng
Water
Up to 14.5 μl
5Â TdT buffer
4 μl
(9.5 mM dCTP + 0.5 mM ddCTP) mix
1 μl
TdT (30 U/μl)
0.5 μl
Total
20 μl
Table 4
PCR mixes for the first PCR round
Reagent
One reaction
Library reaction
Control reaction
Master mix, 2Â
12.5 μl
12.5 μl
Primer 1, 30 μM (TnkN3)
0.5 μl
–
Primer 2, 30 μM (olj376)
1.5 μl
1.5
Water
7.5 μl
8 μl
C-tailed DNA
3 μl
3 μl
Total
25 μl
2 5 μl
Table 5
PCR mixes for the second PCR round
Reagent
One reaction
Library reaction
Control reaction
Master mix, 2Â
25 μl
12.5 μl
Primer 1, 30 μM (pMargent2)
0.5 μl
0.25 μl
Primer 2, 30 μM (indexing primer)
0.5 μl
0.25 μl
Water
23 μl
11.5 μl
PCR products from PCR #1
1 μl
0.5 μl
Total
50 μl
2 5 μl
Transposon Sequencing in Leptospira interrogans
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