...
Add 1 ul of EcoR1 Adaptors with 96-channel and Track which template plate gets which MID plate.
Template | GTL Plate | EcoR1 MID plate | Pool Plate | Library | Norm to | Sequencer |
|---|---|---|---|---|---|---|
WCR31 | 1 | 1+2 | 1 | 10 | Next | |
WCR32 | 2 | 1+2 | 1 | 10 | Next | |
WCR33 | 3 | 3 | 1 | 10 | Next | |
WCR34 | 4 | 4+5 | 2 | 30 | Next | |
WCR36 | 5 | 4+5 | 2 | 30 | Next | |
WCR37 | 6 | 6+7 | 2 | 30 | Next | |
WCR38 | 7 | 6+7 | 3 | 30 | Next | |
WCR39 | 8 | 8+9 | 3 | 30 | Next | |
WCR40 | 1 | 8+9 | 3 | 30 | Next | |
WCR41 | 2 | 10+11 | 4 | 10 | Next | |
WCR45 | 3 | 10+11 | 4 | 10 | Next | |
WCR46 | 4 | 12 | 4 | 10 | Next | |
WCR47 | 5 | 13+14 | 6 | 30 | Nov | |
WCR48 | 6 | 13+14 | 6 | 30 | Nov | |
WCR49 | 7 | 15+16 | 5 | 40 | Nov | |
WCR50 | 8 | 15+16 | 5 | 40 | Nov | |
WCR51 | 1 | 17+18 | 5 | 40 | Nov | |
WCR52 | 2 | 19+20 | 6 | 30 | Nov | |
WCR53 | 3 | 17+18 | 5 | 40 | Nov | |
WCR54 | 4 | 19+20 | 6 | 30 | Nov | |
WCR43 | 5 | 21+22 | 7 | 30 | Nov | |
WCR44 | 6 | 21+22 | 7 | 30 | Nov | |
WCR55 | 7 | 23+24 | 7 | 30 | Nov | |
WCR56 | 8 | 23+24 | 7 | 30 | Nov | |
WCR57 | 1 | 25+26 | 8 | 30 | Next | |
WCR58 | 2 | 25+26 | 8 | 30 | Next | |
WCR59 | 3 | 27+28 | 8 | 30 | Next | |
WCR28 | 47 | 27+28 | 9 | 10* | Next | |
WCR35 | 58 | 29+30 | 9 | 10* | Next | |
WCR42 | 61 | 29+30 | 9 | 10* | Next |
Label reaction plates with MID plate used.
...
Reagent | ul/rxn | rxns | ul needed (x1.4) | |
|---|---|---|---|---|
H2O | 9.52 | 350 | 3094 | 1332.84760 |
5x iProof buffer | 4 | 350 | 1300 | 5602000 |
10 mM dNTPs | 0.4 | 350 | 130 | 56200 |
50 mM MgCl2 | 0.4 | 350 | 130 | 56200 |
5 uM Illumina Primers | 1.33 | 350 | 432 | 186.2665 |
iProof TAQ | 0.2 | 350 | 65 | 28100 |
DMSO | 0.15 | 350 | 49 | 2175 |
total | 16 | 350 | 5200 | 22408000 |
Add 16 ul of PCR1 MM to each well of four new hard shell PCR plates with 8 channel
...
Reagent | ul/rxn | rxns | ul needed (x 1.6) | ||
5x Iproof buffer | 0.425 | 360 | 153 | 65106 | 212.5 |
10 mM dNTPs | 0.4 | 360 | 144 | 61.6100 | 200 |
Primers | 1.33 | 360 | 479 | 332 | 204.8665 |
Total | 2.155 | 360 | 776 | 331.87539 |
Then continue with the last four unlisted steps for GBS1 from above:
...
Run Final Product on qPCR for check and quant
Result from qPCR check :
qPCR | Sample | RSB |
|---|---|---|
2.87 | ||
load =800 pM
dilute 1 nM PhiX to 500 pM
...