CALeDNA_template_amplicon_pooling
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ABCDEFGHIJKLMNOPQRSTUVWXYZAAABACADAEAFAGAHAIAJAKALAMAN
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Amplicon Pooling Template
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1. Indicate the number of samples you have
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AG notes: Do we want option were user can choose to use <10uL of sample?
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Number of samples
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Include in table 1 or 2?
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Double check which calculations are affected
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2. Select the primers you used
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TRUE16s31016s1101016s
16s
310
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TRUE18s27018s2111118s
18s
270
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TRUEFITS370FITS31212FITS
FITS
370
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TRUEPITS520PITS41313PITS
PITS
520
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TRUECO1430CO151414CO1
CO1
430
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FALSEtrnL#REF!
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FALSEOther#REF!
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3. Indicate the PCR Product length and the max volume of sample you wish to use.
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You can batch edit in Table 1, or modify on case by case basis in Table 2. You only need to modify/ verify cells highlighted in blue
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Table 1. Batch editing
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PrimerPCR Product LengthMax volume of sample you would like to add **(considering you eluted to a certain amount, subtracted 2 for quantification and should account for pippeting error- generally recommend 10)
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16s31010
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18s27010
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FITS37010
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PITS52010
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CO143010
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4. Add the concentrations calculated from the Qubit fluorescence to Table 2 below and verify the average amplicon length you observed in your gel.
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5. How much DNA do you want to have in your indexing PCR (generally 5 or 10ng)
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DNA for indexing PCR10ng
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5. Look at results in the "Data output" tab.
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Table 2. Detailed table
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16s18sFITSPITSCO1
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Sample #Enter your sample names herePCR Product LengthConcentration (ng/uL)Max volume of sample you would like to add (uL)PCR Product LengthConcentration (ng/uL)Max volume of sample you would like to add (uL)PCR Product LengthConcentration (ng/uL)Max volume of sample you would like to add (uL)PCR Product LengthConcentration (ng/uL)Max volume of sample you would like to add (uL)PCR Product LengthConcentration (ng/uL)Max volume of sample you would like to add (uL)
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(Example)K0000_A1
16s
3101.463810
18s
2702.963810
FITS
3701.499810
PITS
5202.473810
CO1
4304.763210
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1
16s
31010
18s
27010
FITS
37010
PITS
52010
CO1
43010
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16s
31010
18s
27010
FITS
37010
PITS
52010
CO1
43010
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3
16s
31010
18s
27010
FITS
37010
PITS
52010
CO1
43010
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4
16s
31010
18s
27010
FITS
37010
PITS
52010
CO1
43010
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5
16s
31010
18s
27010
FITS
37010
PITS
52010
CO1
43010
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