Male < 35 FAM > 35 Presence/Absence Female > 35 FAM > 35 Presence/Absence

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1 Page 1 f 5 Please refer t fr updated prtcls and Material Safety Data Sheets (MSDS). Cnsult MSDS befre using any new prduct. FEMINDICATOR is a registered trademark f Medicinal Genmics Crpratin, and is fr labratry use nly. Table f Cntents FemINDICAtr Analysis Quick Table... 1 Detailed Assay Data Analysis... 2 Trubleshting Guide... 4 Glssary and Definitins... 5 LIMITED USE LABEL LICENSE... 5 FemINDICAtR Analysis Quick Table FemINDICAtr Assay Cq Value Flur Negative Cntrl (Cq) Cq threshld Male < 35 FAM > 35 Presence/Absence Female > 35 FAM > 35 Presence/Absence Internal Cntrl* <35 HEX *Internal cntrl verifies the presence r absence f plant DNA Assay Psitive Cntrls <35 FAM

2 Page 2 f 5 Detailed Assay Data Analysis 1. Presence / Absence Assays 1.1. The Data Analysis windw will pen autmatically when the run is cmplete Highlight the well f interest. The graph will appear abve. The Cq values will appear t the right T analyze the results Start by turning the graph t Lg Scale and manually mving the threshld t 10 2 fr the FAM flurphre. Als manually mve the threshld fr the HEX flurphre half way between 10 2 and T turn the graph t Lg Scale, click n the bx at the bttm right f the graph. T mve the threshld, click n the hrizntal lines, and mve them t 10 2 n the y-axis. Threshld Line Lg Scale Cntrls Male specific Psitive Cntrl, n the FAM flurphre, has a Cq value < 35. Visually cnfirm with the curve n the graph. Male specific Negative Cntrl, n the FAM flurphre, has a Cq value > 35 r n Cq value. Visually cnfirm with the curve n the graph. Unknwn Samples Internal Cntrl, n the HEX flurphre, has a Cq value < 35. Visually cnfirm with the curve n the graph A presence r male result fr the unknwn sample. Any Cq value fr the FAM flurphre < 35. Visually cnfirm with the curve n the graph. (It is very imprtant t cnfirm with the amplificatin curve when a presence result ccurred. Smetimes the backgrund amplificatin will give a false psitive reading, especially when Cq reading is less than 15. See trubleshting guide belw.) An absence r female result fr the unknwn sample. N Cq value fr the FAM flurphre r a Cq value >35. Visually cnfirm n curve n the graph.

3 Page 3 f 5 2. Exprt the Data 2.1. Exprting the Cq values int an Excel spreadsheet. T exprt the Cq values t an Excel spreadsheet, right- click n the chart n the bttm right f the screen. Chse Exprt T Excel 2.2. Saving a visual f the graph T save a picture f the graph, right click n the graph n the tp left f the screen. Chse Save Image As

4 Page 4 f 5 Trubleshting Guide Symptm Reasn Slutin Extractin Failure Repeat SenSATIVAx and FemINDICAtr by Residual ethanl in elutin Ethanl is an inhibitr t PCR. Return t the SenSATIVAx TM prtcl and repeat all steps. Mix up in Reactin Repeat the Decntaminatin and qpcr by Internal cntrl Setup (SCCG Primer) failure In the Data Analysis windw click n View/Edit Missing Flurphre n plate set up Plate Setup frm the Settings drp dwn. All wells shuld have bth FAM and HEX. Once cmpleted and windw is clsed the analysis shuld autmatically update. Visually cnfirm that there is an amplificatin Small Cq value <15 curve. If nt, this is lw level backgrund and is t be expected. Psitive Negative Cntrl Carry ver Repeat the Decntaminatin and qpcr by Insufficient presetup bleaching Wipe dwn the lab wrkspace and all equipment with 10% Bleach. Repeat Decntaminatin and qpcr Prtcls. Negative Psitive Cntrl Mix up in Reactin Setup Repeat the Decntaminatin and qpcr by Ttal run failure Excessive vrtex f the qpcr Master Mix Repeat the Decntaminatin and qpcr by Backgrund Amplificatin Unclear This is usually seen with a very lw Cq reading (<15), the curve is usually missing the expnential grwth phase, but rather a gradual increase f flrescent signal. This is usually a negative result, but shuld be repeated.

5 Page 5 f 5 Glssary and Definitins Dexyribnucleic acid (DNA) is a mlecule that encdes the genetic instructins used in the develpment and functining f all knwn living rganisms. Plymerase Chain Reactin (PCR) is a technlgy in mlecular bilgy used t amplify a single cpy r a few cpies f a piece f DNA acrss several rders f magnitude, generating thusands t millins f cpies f a particular DNA sequence. A flurphre is a flurescent chemical cmpund that can re-emit light upn light excitatin. The Negative Cntrls are the samples where n r a very high Cq is expected. It helps t ensure that all Assay specific reactins are clean f cntaminates. The assay specific Psitive Cntrls are the samples where a Cq is expected. It helps ensure that all Assay specific reactins are wrking crrectly. The Assay specific Psitive Cntrl is targeting the pathgen using the FAM flurphre. The Internal Cntrl is added t every sample where a Cq is expected. It ensures the effectiveness and efficiency f each reactin. The internal cntrl is targeting a Single Cpy Cntrl Gene r SCCG, using the HEX flurphre. LEGAL DISLCAIMER This test was develped as part f a research prject cnducted by Medicinal Genmics Crpratin ( MGC ). Neither MGC nr any f their emplyees, cntractrs r ther affiliates, makes any warranty, express r implied, r assumes any legal liability r respnsibility fr the accuracy, cmpleteness, r any third party's use r the results f such use f any infrmatin LIMITED USE LABEL LICENSE This prduct is cvered by at least ne r mre claims f US patents applicatins, which are exclusively licensed t Medicinal Genmics Crpratin. This prduct is sld strictly fr the use f the buyer, and the buyer is nt authrized t transfer this prduct [r any materials made using this prduct] t any third party Medicinal Genmics Crpratin. All rights reserved. * All Trademarks are prperty f their respective wners.