Listeria Quick Guide - Bio-Rad Laboratories · Bio-Rad, S.N.C. au capital de 50 000 000 Euros,...
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Please read the kit instruction manual and instrument user guide for complete and detailed instructions. • Enrich the sample in LSB (25 g in 225 ml), 25 hrs ± 1 hr at 30°C • Add 100 μl of the complete lysis reagent (reagent A + reagent F) in the deepwell plate Lysis reagent must be constantly stirring in order to keep it in suspension Use a micropipette with large opening 200 μl filter tips • Transfer 100 μl of enriched sample Avoid including large fragments of food debris, and shaking stomacher bag before collecting • Seal the deepwell plate with the pre-pierced sealing film • Incubate at 95-100°C for 15-20 min at 1,300 rpm in a plate agitator- incubator • Cool the deepwell plate • Start software • Create the plate setup • Start the amplification by clicking on “Run” 25 hrs ± 1 hr 30°C LSB 357-8113 • iQ-Check ™ Listeria spp. Easy Extraction Deepwell Protocol Quick Guide 100 μl 100 μl 15-20 min 1,300 rpm 95-100°C • Prepare the PCR mix • Distribute 45 μl/well in the PCR microplate probe mix amplification mix PCR mix 45 μl • Add 5 μl of controls • Transfer 5 μl of the sample supernatants Do not vortex before collecting the sample Check there are no bubbles • Seal the microplate 5 μl
Transcript of Listeria Quick Guide - Bio-Rad Laboratories · Bio-Rad, S.N.C. au capital de 50 000 000 Euros,...
Please read the kit instruction manual and instrument user guide for complete and detailed instructions.
• Enrich the sample in LSB (25 g in 225 ml), 25 hrs ± 1hr at 30°C
• Add 100 μl of the complete lysis reagent (reagent A + reagent F) in thedeepwell plateLysis reagent must be constantly stirring in order to keep it in suspensionUse a micropipette with large opening 200 μl filter tips
• Transfer 100 μl of enriched sampleAvoid including large fragments of food debris, and shaking stomacherbag before collecting
• Seal the deepwell plate with the pre-pierced sealing film
• Incubate at 95-100°C for 15-20 min at 1,300 rpm in a plate agitator-incubator
• Cool the deepwell plate
• Start software
• Create the plate setup
• Start the amplification by clicking on “Run”
25 hrs± 1 hr
30°CLSB
357-8113 • iQ-Check™ Listeria spp.Easy ExtractionDeepwell Protocol
Quick Guide
100 μl 100 μl
15-20 min1,300 rpm
95-100°C
• Prepare the PCR mix• Distribute 45 μl/well in the PCR microplate
probemix
amplificationmix
PCRmix
45 μl
• Add 5 μl of controls
• Transfer 5 μl of the sample supernatantsDo not vortex before collecting the sampleCheck there are no bubbles
• Seal the microplate
5 μl
iQ-Check™ Listeria spp.PCR Mix Calculation Guide
Quick Guide
To find the correct volumes to use when preparing the PCR mix, add the total number of samples and controls tobe analyzed, and find the corresponding volumes of reagent B and reagent C in the table.
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© Bio-Rad PF098 Rev C 07/2011
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Total number ofsamples & controls
Probes -Reagent B (μl)
Amplification MixReagent C (μl)
1 5 402 11 863 16 1304 22 1735 27 2166 32 2597 38 3028 43 3469 49 38910 54 43211 59 47512 65 51813 70 56214 76 60515 81 64816 86 69117 92 73418 97 77819 103 82120 108 86421 113 90722 119 95023 124 99424 130 100025 135 110026 140 110027 146 120028 151 120029 157 130030 162 130031 167 130032 173 140033 178 140034 184 150035 189 150036 194 160037 200 160038 205 160039 211 170040 216 170041 221 180042 227 180043 232 190044 238 190045 243 190046 248 200047 254 200048 259 2100
Total number ofsamples & controls
Probes -Reagent B (μl)
Amplification MixReagent C (μl)
49 265 210050 270 220051 275 220052 281 220053 286 230054 292 230055 297 240056 302 240057 308 250058 313 250059 319 250060 324 260061 329 260062 335 270063 340 270064 346 280065 351 280066 356 290067 362 290068 367 290069 373 300070 378 300071 383 310072 389 310073 394 320074 400 320075 405 320076 410 330077 416 330078 421 340079 427 340080 432 350081 437 350082 443 350083 448 360084 454 360085 459 370086 464 370087 470 380088 475 380089 481 380090 486 390091 491 390092 497 400093 502 400094 508 410095 513 410096 518 4100
