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=Overnight Capture (Started [[Daniel:Notebook/ComboLock/2016-12-15|Yesterday]])= [[Daniel:Notebook/ComboLock|Back to Calendar]] ==Protocol== <ol start="4"> <li>Circularization</li> <ol type="A"> <li>Prepare 7X uL Phusion mix</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#C4D79B;font-size:12pt;font-weight:bold" align="center" | width="150" height="30" | Reagent | width="120" | Stock Conc | width="74" | Final Amount | width="65" | 1x Vol (uL) | width="90" | MM Vol (7x) (uL) |- style="font-size:12pt" | height="15" valign="bottom" | NAD+ | align="center" | 5 mM | align="center" | 40 nmol | align="center" align="center" | 8 | align="center" align="center" | 56 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | dNTP | align="center" | 1 mM | align="center" | 600 pmol | align="center" align="center" | 0.6 | align="center" align="center" | 4.2 |- style="font-size:12pt" | height="15" valign="bottom" | Betaine | align="center" | 5 M | align="center" | 15 umol | align="center" align="center" | 3 | align="center" align="center" | 21 |- style="background-color:#BFBFBF;font-size:12pt" | height="30" valign="bottom" | 10X AmpLigase Buffer | align="center" | 10X | align="center" | 1X | align="center" align="center" | 2 | align="center" align="center" | 14 |- style="font-size:12pt" | height="15" valign="bottom" | Amp Ligase | align="center" | 5 U/uL | align="center" | 10 U | align="center" align="center" | 2 | align="center" align="center" | 14 |- style="background-color:#BFBFBF;font-size:12pt" | height="30" valign="bottom" | Phusion HF DNA Polymerase | align="center" | 2000 U/mL | align="center" | 6.4U | align="center" align="center" | 3.2 | align="center" align="center" | 22.4 |- style="font-size:12pt" | height="15" valign="bottom" | nf H2O | align="center" align="center" | | align="center" align="center" | | align="center" align="center" | 1.2 | align="center" align="center" | 8.4 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | Total | align="center" align="center" | | align="center" align="center" | | align="center" align="center" | 20 | align="center" align="center" | 140 |} <li>Add 20 uL Phusion mix to 20 uL of reaction</li> <li>Incubate at 60C for 2 hours</li></ol> <li>Exonuclease Digestion</li> <ol type="A"> <li>Mix 12.5 uL Exo I (20U/uL) and 2.5 uL ExoIII (100U/uL)</li> <li>Add 2 uL to each sample, mix by swirling pipette tip</li> <li>Incubate for 1 hour at 37C</li> <li>Heat kill by incubating for 5 min at 95C</li> </ol> <li>Rolling Circle Amplification</li> <ol type="A"> <li>Prepare 7X Master mix according to table below</li> <li>Add 15 uL master mix to each tube</li> <li>Add sample to new reaction tubes according to following table; '''use template from both supernatant only and beads'''</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="160" height="30" | Reagent | width="85" | uL Added | width="85" | Master Mix (13X) |- style="font-size:12pt" | height="15" valign="bottom" | Template | align="center" align="center" valign="bottom" | 5 | align="center" valign="bottom" | NA |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | RCA Primer (100 uM) | align="center" align="center" valign="bottom" | 2.5 | align="center" align="center" valign="bottom" | 32.5 |- style="font-size:12pt" | height="15" valign="bottom" | dNTP (1 mM) | align="center" align="center" valign="bottom" | 0.8 | align="center" align="center" valign="bottom" | 10.4 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | 10X Buffer | align="center" align="center" valign="bottom" | 2 | align="center" align="center" valign="bottom" | 26 |- style="font-size:12pt" | height="15" valign="bottom" | Phi29 | align="center" align="center" valign="bottom" | 1 | align="center" align="center" valign="bottom" | 13 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | nfH2O | align="center" align="center" valign="bottom" | 8.7 | align="center" align="center" valign="bottom" | 113.1 |- style="font-size:12pt" | height="15" valign="bottom" | Total |style="font-weight:bold" align="center" align="center" valign="bottom" | 20 |style="font-weight:bold" align="center" align="center" valign="bottom" | 195 |} <li>Incubate at 37C for 3 hours</li> <li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol> <li>qPCR</li> <ol type="A"> <li>Make master mix according to following recipe</li> <ol type="a"> <li>151.2 uL nfH2O</li> <li>180 uL SYBR Fast</li> <li>7.2 uL 10 uM AmpF</li> </ol> <li>Add 47 uL master mix to each well</li> <li>Add 2 uL sample and 1 uL appropriate Reverse Primer (1 uM) according to table</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="90" height="45" | Sample | width="105" | Condition | width="105" | Fraction | width="55" | Lane | width="49" | AmpR Index | width="53" | Sample Vol (uL) | width="65" | 2X Kapa SYBR qPCR MM | width="65" | 10 uM Forward Primer | width="65" | 10 uM Reverse Primer | width="65" | H2O | width="65" | Total Volume (uL) |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 1A | align="center" | Sample | align="center" | Supernatant | align="center" | A1 | align="center" align="center" | 20 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="15" | Sample 1B | align="center" | Sample | align="center" | Supernatant | align="center" | A2 | align="center" align="center" | 20 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 2A | align="center" | No Template | align="center" | Supernatant | align="center" | A3 | align="center" align="center" | 21 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | Sample 2B | align="center" valign="bottom" | No Template | align="center" | Supernatant | align="center" | A4 | align="center" align="center" | 21 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 3A | align="center" | No C Probe | align="center" | Supernatant | align="center" | A5 | align="center" align="center" | 22 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="15" | Sample 3B | align="center" | No C Probe | align="center" | Supernatant | align="center" | A6 | align="center" align="center" | 22 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 1A | align="center" | Sample | align="center" | Beads | align="center" | A1 | align="center" align="center" | 20 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" | Sample 1B | align="center" | Sample | align="center" | Beads | align="center" | A2 | align="center" align="center" | 20 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 2A | align="center" | No Template | align="center" | Beads | align="center" | A3 | align="center" align="center" | 21 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | Sample 2B | align="center" valign="bottom" | No Template | align="center" | Beads | align="center" | A4 | align="center" align="center" | 21 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 3A | align="center" | No C Probe | align="center" | Beads | align="center" | A5 | align="center" align="center" | 22 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" | Sample 3B | align="center" | No C Probe | align="center" | Beads | align="center" | A6 | align="center" align="center" | 22 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 |- style="font-size:12pt" | height="15" valign="bottom" | NTC | align="center" valign="bottom" | qPCR Primers Only | align="center" align="center" valign="bottom" | | align="center" | A7 | align="center" align="center" | 23 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 23 | align="center" align="center" | 50 |} </ol> ===Results=== <gallery perrow=2 heights=300px widths=300px> File:|qPCR curve File:2016-12-16-MirrorCProbe-ONCap-Supernatant.png|Gel image-supernatant RCA File:2016-12-16-MirrorCProbe-ONCap-Beads.png|Gel image-Beads RCA </gallery> ==Adapter Testing== Just like [[Daniel:Notebook/ComboLock/2016-12-15|yesterday]] I'm going to test presence of C probes and circular product in the pre-RCA mix. <ol start="7"> <li>qPCR</li> <ol type="A"> <li>Make master mix according to following recipes</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | width="85" height="45" | Master Mix | width="85" | Primers | width="65" | Primer Target | width="85" | 2X Kapa SYBR qPCR MM | width="85" | 10 uM Forward Primer | width="85" | 10 uM Reverse Primer | width="65" | H2O | width="65" | Master Mix |- style="font-size:12pt" |style="font-weight:bold" height="15" | MMC1 | align="center" | p24/p4* | align="center" | C1 | align="center" align="center" | 230 | align="center" align="center" | 9.2 | align="center" align="center" | 9.2 | align="center" align="center" | 193.2 | align="center" | MMC1 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="15" | MMC2 | align="center" | p24/p4* | align="center" | C1 | align="center" align="center" | 230 | align="center" align="center" | 9.2 | align="center" align="center" | 9.2 | align="center" align="center" | 193.2 | align="center" | MMC1 |- style="font-size:12pt" |style="font-weight:bold" height="15" | MMCP | align="center" | p6/p12* | align="center" | C2 | align="center" align="center" | 230 | align="center" align="center" | 9.2 | align="center" align="center" | 9.2 | align="center" align="center" | 193.2 | align="center" | MMC2 |} <li>Add 48 uL master mix to each well</li> <li>Add 2 uL sample according to table in the following section</li> <li>qPCR Cycles</li> <ol type="a"> <li>95C 3 min</li> <li>95C 3 sec</li> <li>55C 30 sec</li> <li>72C 20 sec</li> <li>plate read</li> <li>goto b x24</li> <li>72C 2 min</li> <li>16C hold</li> </ol></ol> <li>TBE Gel</li> <ol type="A"> <li>Mix 144 uL TBE, 36 uL 6x loading dye</li> <li>Aliquot 10 uL per sample/ladder lane onto parafilm</li> <li>Add 2 uL of sample or ladder to correct drop</li> <li>Load 10 uL in to well</li> <li>Run gel for 22 minutes at 250V</li> <li>Open gel and stain with 3 uL SYBR Gold for 3 minutes</li> <li>Rinse gel and image in gel doc</li> </ol></ol> ===Results=== <gallery perrow=3 heights=300px widths=300px> File:|qPCR curve displaying sample vs C probes (for supernatant) File:|qPCR curve displaying sample vs C probes (for beads) File:|qPCR curve displaying supernatant vs. beads effect File:2016-12-16-MirrorCProbe-ONCap-Adaptors-CProbes.png|Gel image 1-Sample C probe detection File:2016-12-16-MirrorCProbe-ONCap-Adaptors-Controls.png|Gel image 2-Controls gel </gallery> ===qPCR Sample Matrix=== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="100" height="45" | Sample | width="120" | Condition | width="90" | Primers | width="130" | Primer Target | width="91" | Fraction | width="65" | Lane | width="65" | Sample Vol (uL) | width="73" | 2X Kapa SYBR qPCR MM | width="80" | 10 uM Forward Primer | width="80" | 10 uM Reverse Primer | width="65" | H2O | width="65" | Total Volume (uL) | width="65" | Master Mix |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 1A | align="center" | Sample | align="center" | p24/p4* | align="center" | C1 | align="center" | Supernatant-PreRCA | align="center" | A1 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="30" | Sample 1B | align="center" | Sample | align="center" | p24/p4* | align="center" | C1 | align="center" | Supernatant-PreRCA | align="center" | A2 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 1A | align="center" | Sample | align="center" | p6/p12* | align="center" | C2 | align="center" | Supernatant-PreRCA | align="center" | A3 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="30" | Sample 1B | align="center" | Sample | align="center" | p6/p12* | align="center" | C2 | align="center" | Supernatant-PreRCA | align="center" | A4 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 1A | align="center" | Sample | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Supernatant-PreRCA | align="center" | A5 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="background-color:#BFBFBF;font-size:12pt" |style="font-weight:bold" height="30" | Sample 1B | align="center" | Sample | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Supernatant-PreRCA | align="center" | A6 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 1A | align="center" | Sample | align="center" | p24/p4* | align="center" | C1 | align="center" | Beads-PreRCA | align="center" | C1 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" | Sample 1B | align="center" | Sample | align="center" | p24/p4* | align="center" | C1 | align="center" | Beads-PreRCA | align="center" | C2 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 1A | align="center" | Sample | align="center" | p6/p12* | align="center" | C2 | align="center" | Beads-PreRCA | align="center" | C3 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" | Sample 1B | align="center" | Sample | align="center" | p6/p12* | align="center" | C2 | align="center" | Beads-PreRCA | align="center" | C4 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 1A | align="center" | Sample | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Beads-PreRCA | align="center" | C5 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="background-color:#DA9694;font-size:12pt" |style="font-weight:bold" height="15" | Sample 1B | align="center" | Sample | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Beads-PreRCA | align="center" | C6 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 3A | align="center" | No C Probes | align="center" | p24/p4* | align="center" | C1 | align="center" | Supernatant-PreRCA | align="center" | F1 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="background-color:#C4D79B;font-size:12pt" |style="font-weight:bold" height="30" | Sample 3B | align="center" | No C Probes | align="center" | p24/p4* | align="center" | C1 | align="center" | Supernatant-PreRCA | align="center" | F2 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 3A | align="center" | No C Probes | align="center" | p6/p12* | align="center" | C2 | align="center" | Supernatant-PreRCA | align="center" | F3 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="background-color:#C4D79B;font-size:12pt" |style="font-weight:bold" height="30" | Sample 3B | align="center" | No C Probes | align="center" | p6/p12* | align="center" | C2 | align="center" | Supernatant-PreRCA | align="center" | F4 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="font-size:12pt" |style="font-weight:bold" height="30" | Sample 3A | align="center" | No C Probes | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Supernatant-PreRCA | align="center" | F5 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="background-color:#C4D79B;font-size:12pt" |style="font-weight:bold" height="30" | Sample 3B | align="center" | No C Probes | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Supernatant-PreRCA | align="center" | F6 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 3A | align="center" | No C Probes | align="center" | p24/p4* | align="center" | C1 | align="center" | Beads-PreRCA | align="center" | H1 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="background-color:#B1A0C7;font-size:12pt" |style="font-weight:bold" height="15" | Sample 3B | align="center" | No C Probes | align="center" | p24/p4* | align="center" | C1 | align="center" | Beads-PreRCA | align="center" | H2 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC1 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 3A | align="center" | No C Probes | align="center" | p6/p12* | align="center" | C2 | align="center" | Beads-PreRCA | align="center" | H3 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="background-color:#B1A0C7;font-size:12pt" |style="font-weight:bold" height="15" | Sample 3B | align="center" | No C Probes | align="center" | p6/p12* | align="center" | C2 | align="center" | Beads-PreRCA | align="center" | H4 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMC2 |- style="font-size:12pt" |style="font-weight:bold" height="15" | Sample 3A | align="center" | No C Probes | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Beads-PreRCA | align="center" | H5 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="background-color:#B1A0C7;font-size:12pt" |style="font-weight:bold" height="15" | Sample 3B | align="center" | No C Probes | align="center" | p24/p12* | align="center" | Circular Product | align="center" | Beads-PreRCA | align="center" | H6 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 21 | align="center" align="center" | 50 | align="center" | MMCP |- style="font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | NTC | align="center" valign="bottom" | qPCR Primers Only | align="center" valign="bottom" | p24/p4* | align="center" valign="bottom" | C1 | align="center" valign="bottom" | NA | align="center" | A7 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 23 | align="center" align="center" | 50 | align="center" | MMC1 |- style="background-color:#FABF8F;font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | NTC | align="center" valign="bottom" | qPCR Primers Only | align="center" valign="bottom" | p6/p12* | align="center" valign="bottom" | C2 | align="center" valign="bottom" | NA | align="center" | C7 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 23 | align="center" align="center" | 50 | align="center" | MMC2 |- style="font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | NTC | align="center" valign="bottom" | qPCR Primers Only | align="center" valign="bottom" | p24/p12* | align="center" valign="bottom" | Circular Product | align="center" valign="bottom" | NA | align="center" | F7 | align="center" align="center" | 2 | align="center" align="center" | 25 | align="center" align="center" | 1 | align="center" align="center" | 1 | align="center" align="center" | 23 | align="center" align="center" | 50 | align="center" | MMCP |} [[Category:ComboLock]] [[Category:20161215]]
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