A | B | C | |
Oligo ID | Sequence (5’ → 3’) | Purification / synthesis scale | |
Smart dT30VN | /5Biosg/AAGCAGTGGTATCAACGCAGAGTACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN | desalted or HPLC | |
FS TSO | /5Biosg/AAGCAGTGGTATCAACGCAGAGTACrGrGrG | desalted or HPLC |
A | B | C | |
Oligo ID | Sequence (5’ → 3’) | Purification / synthesis scale | |
TN5MErev | /5Phos/ CTGTCTCTTATACACATCT | 2 µM scale - desalted* | |
TN5ME-A | TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG | 1 µM scale - desalted* | |
TN5ME-B | GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG | 1 µM scale - desalted* |
A | B | |
Oligo ID | Sequence (5’ → 3’) | |
Nextera_v2_N714 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCATGAGCGTCTCGTGGGCTCG*G | |
Nextera_v2_N715 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCCTGAGATGTCTCGTGGGCTCG*G | |
Nextera_v2_N716 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTAGCGAGTGTCTCGTGGGCTCG*G | |
Nextera_v2_N718 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTAGCTCCGTCTCGTGGGCTCG*G | |
Nextera_v2_N719 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTACTACGCGTCTCGTGGGCTCG*G | |
Nextera_v2_N720 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAGGCTCCGGTCTCGTGGGCTCG*G | |
Nextera_v2_N721 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGCAGCGTAGTCTCGTGGGCTCG*G | |
Nextera_v2_N722 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTGCGCATGTCTCGTGGGCTCG*G | |
Nextera_v2_N723 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGAGCGCTAGTCTCGTGGGCTCG*G | |
Nextera_v2_N724 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCGCTCAGTGTCTCGTGGGCTCG*G | |
Nextera_v2_N726 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTCTTAGGGTCTCGTGGGCTCG*G | |
Nextera_v2_N727 | /5Biosg/CAAGCAGAAGACGGCATACGAGATACTGATCGGTCTCGTGGGCTCG*G | |
Nextera_v2_N728 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTAGCTGCAGTCTCGTGGGCTCG*G | |
Nextera_v2_N729 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGACGTCGAGTCTCGTGGGCTCG*G | |
Nextera_v2_S502 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCTCTCTATTCGTCGGCAGCGT*C | |
Nextera_v2_S513 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTCGACTAGTCGTCGGCAGCGT*C | |
Nextera_v2_S503 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTATCCTCTTCGTCGGCAGCGT*C | |
Nextera_v2_S515 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTTCTAGCTTCGTCGGCAGCGT*C | |
Nextera_v2_S505 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGTAAGGAGTCGTCGGCAGCGT*C | |
Nextera_v2_S516 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCCTAGAGTTCGTCGGCAGCGT*C | |
Nextera_v2_S506 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACACTGCATATCGTCGGCAGCGT*C | |
Nextera_v2_S517 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGCGTAAGATCGTCGGCAGCGT*C | |
Nextera_v2_S507 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACAAGGAGTATCGTCGGCAGCGT*C | |
Nextera_v2_S518 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCTATTAAGTCGTCGGCAGCGT*C | |
Nextera_v2_S508 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCTAAGCCTTCGTCGGCAGCGT*C | |
Nextera_v2_S520 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACAAGGCTATTCGTCGGCAGCGT*C | |
Nextera_v2_S510 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCGTCTAATTCGTCGGCAGCGT*C | |
Nextera_v2_S521 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGAGCCTTATCGTCGGCAGCGT*C | |
Nextera_v2_S511 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTCTCTCCGTCGTCGGCAGCGT*C | |
Nextera_v2_S522 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTTATGCGATCGTCGGCAGCGT*C |
A | B | |
Oligo ID | Sequence | |
Nextera_extra_i7_1 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGCCTATCAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_2 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTTGGATGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_3 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAGTCTCACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_4 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTCATCAGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_5 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGTACCGTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_6 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAAGTCGAGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_7 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCACGTTGTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_8 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCACAGCAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_9 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTACTTGGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_10 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCCTCAGTTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_11 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCCTACCTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_12 | /5Biosg/CAAGCAGAAGACGGCATACGAGATATGGCGAAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_13 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTTACCTGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_14 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTCGATACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_15 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCCGTGAAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_16 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTAGAGCTCGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_17 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGACTGACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_18 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTAGACGTGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_19 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCCGGAATTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_20 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTCCTAGAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_21 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCAACGGATGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_22 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGGCTATCGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_23 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCGGTCATAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_24 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCCAATCGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_25 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGAGCTTGTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_26 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGAAGGTTCGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_27 | /5Biosg/CAAGCAGAAGACGGCATACGAGATATCTCGCTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_28 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAGTTACGGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_29 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTGTCTGAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_30 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGACTTCGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_31 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGGATCACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_32 | /5Biosg/CAAGCAGAAGACGGCATACGAGATACACCAGTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_33 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCAGGTTAGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_34 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAGTTGGCTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_35 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCAACTGGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_36 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCTGCACTTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_37 | /5Biosg/CAAGCAGAAGACGGCATACGAGATACACGGTTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_38 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAATACGCGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_39 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGCGAACTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_40 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGCTGACTAGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_41 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTGGTGTTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_42 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTGCTTACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_43 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTCAAGGACGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_44 | /5Biosg/CAAGCAGAAGACGGCATACGAGATTGAACCTGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_45 | /5Biosg/CAAGCAGAAGACGGCATACGAGATAGTGTTGGGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_46 | /5Biosg/CAAGCAGAAGACGGCATACGAGATGTACTCTCGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_47 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCCGTATCTGTCTCGTGGGCTCG*G | |
Nextera_extra_i7_48 | /5Biosg/CAAGCAGAAGACGGCATACGAGATCGAAGAACGTCTCGTGGGCTCG*G |
A | B | |
Oligo ID | Sequence | |
Nextera_extra_i5_1 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCGACCATTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_2 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGATAGCGATCGTCGGCAGCGT*C | |
Nextera_extra_i5_3 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACAATGGACGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_4 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCGCTAGTATCGTCGGCAGCGT*C | |
Nextera_extra_i5_5 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTCTCTAGGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_6 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACACATTGCGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_7 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTGAGGTGTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_8 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACAATGCCTCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_9 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCTGGAGTATCGTCGGCAGCGT*C | |
Nextera_extra_i5_10 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGTATGCTGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_11 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTGGAGAGTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_12 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCGATAGAGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_13 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCTCATTGCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_14 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACACCAGCTTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_15 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGAATCGTGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_16 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACAGGCTTCTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_17 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCAGTTCTGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_18 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTTGGTGAGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_19 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCATTCGGTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_20 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTGTGAAGCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_21 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTAAGTGGCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_22 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACACGTGATGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_23 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGTAGAGCATCGTCGGCAGCGT*C | |
Nextera_extra_i5_24 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGTCAGTTGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_25 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACATTCGAGGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_26 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGATACTGGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_27 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGCCTTGTTTCGTCGGCAGCGT*C | |
Nextera_extra_i5_28 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACTTGGTCTCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_29 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACCCGACTATTCGTCGGCAGCGT*C | |
Nextera_extra_i5_30 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGTCCTAAGTCGTCGGCAGCGT*C | |
Nextera_extra_i5_31 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACACCAATGCTCGTCGGCAGCGT*C | |
Nextera_extra_i5_32 | /5Biosg/AATGATACGGCGACCACCGAGATCTACACGATGCACTTCGTCGGCAGCGT*C |
A | B | C | D | |
Reagent | Reaction concentration | Volume (µl) | 384-well plate | |
Triton-X100 (10% v/v) | 0.2% | 0.020 | 8.448 | |
dNTP mix (25 mM each) | 6 mM | 0.240 | 101.376 | |
SMART dT30VN (100 µM) | 1.8 µM | 0.018 | 7.603 | |
RNAse inhibitor (40 U/µl) | 1.2 U/µl | 0.030 | 12.672 | |
DTT (100 mM) | 1.2 mM | 0.012 | 5.069 | |
FS TSO (100 µM) | 9.2 µM | 0.092 | 38.861 | |
dCTP (100 mM) | 9 mM | 0.090 | 38.016 | |
Betaine (5 M) | 1 M | 0.200 | 84.480 | |
Nuclease-free water | - | 0.298 | 125.875 | |
Total volume (µl) | 1.000 | 422.400 |
A | B | C | D | |
Reagent | Reaction concentration | Volume (µl) | 384-well plate | |
DTT (0.1 M) | 4.8 mM | 0.238 | 100.531 | |
MgCl2 (1 M) | 9.2 mM | 0.046 | 19.430 | |
Betaine (5 M) | 800 mM | 0.800 | 337.920 | |
RNAse inhibitor (40 U/µl) | 0.8 U/µl | 0.096 | 40.550 | |
SuperScript IV (200 U/µl) | 2.00 U/µl | 0.050 | 21.120 | |
KAPA HiFi HotStart ReadyMix (2 x) | 1 x | 2.500 | 1056.000 | |
Nuclease-free water | - | 0.270 | 114.048 | |
Total volume (µl) | 4.000 | 1689.600 |
A | B | C | D | E | |
Step | Temperature | Time | Cycles | ||
RT | 50ºC | 60 min | 1 x | ||
PCR | initial denaturation | 98ºC | 3 min | 1 x | |
denaturation | 98ºC | 20 sec | 10-16 x* | ||
annealing | 67ºC | 20 sec | |||
elongation | 72ºC | 6 min | |||
15ºC | Hold |
A | B | C | |
Reagent | Volume (µl) | Final concentration | |
TAPS-Mg buffer, pH=7.3 (5x) | 2.000 | 10 mM TAPS, 5 mM MgCl2 | |
Dimethylformamide (DMF) (100%) | 2.000 | 20% | |
Tn5 transposase (2 µM working dil.) | 0.025 | 5 nM | |
Nuclease-free water | 4.975 | ||
Total volume (µl) | 9.000 |
A | B | C | |
Reagent | Volume (µl) | Final concentration | |
KAPA HiFi enzyme (1 U/μl) | 0.50 | 0.02 U/μl | |
KAPA HiFi Buffer (5 x) | 5.00 | 1 x | |
dNTPs (10 mM) | 0.75 | 300 nM | |
Nuclease-free water | 3.75 | ||
Total volume (µl) | 10.00 |
A | B | C | D | E | |
Step | Temperature | Time | Cycles | ||
gap filling | 72ºC | 3 min | 1 x | ||
enrichment PCR | initial denaturation | 98ºC | 30 sec | 1 x | |
denaturation | 98ºC | 10 sec | 14-16 x | ||
annealing | 55ºC | 30 sec | |||
elongation | 72ºC | 30 sec | |||
15ºC | hold |