Retsch MM400 Ball Mill (Laichi co., Berlin, Germany); PTC200PCR (Bio‐Rad, Berkeley, CA, USA); GelDox XR gel imaging system (Bio‐Rad, USA); DYY‐8C Electrophoretic apparatus (Beijing Liuyi Instrument, China); BI3730XL sequenator (Applied Biosystems, Foster City, CA, USA); KRQ‐300P manual climatic box (ChongQing YinHe Test Instrument, China); Plant DNA Extraction Kit (TIANGEN Biotech Co., Chengdu, China); 2 × Taq PCR Master Mix (TIANGEN Biotech Co.,); primer was compounded by Sangon Co., China. Shimadzu; DM4000M microscope (Leica, Frankfurt, Germany); TruSeq® DNA PCR‐Free Sample Preparation Kit (Illumina Co., San Diego, CA, USA); QIAquick gum recovery kit (QIAGEN, Duesseldorf, Germany); Phusion® High‐Fidelity PCR Master Mix with GC Buffer (New England Biolabs, USA); High‐efficient enzyme (New England Biolabs, Boston, MA, USA).
Ptc200pcr
Manufactured by Bio-Rad
Sourced in Germany
The PTC200 Peltier Thermal Cycler is a laboratory instrument designed for performing polymerase chain reaction (PCR) experiments. It is capable of precisely controlling the temperature of samples during the different stages of the PCR process, which is essential for the amplification of DNA or RNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
Plant dna extraction kit, by Tiangen Biotech (2 mentions)
Bi3730xl sequenator, by Thermo Fisher Scientific (2 mentions)
Dm4000m microscope, by Leica (2 mentions)
Geldox xr gel imaging system, by Bio-Rad (2 mentions)
Taq pcr mastermix, by Tiangen Biotech (2 mentions)
Phusion high fidelity pcr master mix with gc buffer, by New England Biolabs (1 mentions)
Massarray typer software version 4, by Labcorp (1 mentions)
Bigdye terminator 3.1 cycle sequencing kit, by Thermo Fisher Scientific (1 mentions)
Abi 3500xl genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Naringenin, by Agilent Technologies (1 mentions)
Lc 20at hplc, by Shimadzu (1 mentions)
4 protocols using ptc200pcr
1
Plant DNA Extraction and Sequencing
2
Genetic Profiling of Tumor Samples
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AS QIAamp DNA FFPE tissue kit (Qiagen, Hilden, Germany) was used to extract DNA from paraffin-embedded tissue samples, and the presence of tumor cells (>70%) was obtained by trimming the normal and necrotic tissues. Genetic analyses of EGFR, KRAS, and BRAF were performed using the OncoCarta Panel14 (link) (version 1.0; Sequenom, San Diego, CA, USA). Mutation data were analyzed using MassArray Typer software version 4.0 (Sequenom). MET skipping mutation was detected by direct sequencing6 (link) using a PTC-200PCR (Bio-Rad, Hercules, CA, USA) with the forward primer 5’-CTTTGTACGTCTCATGTTAT-3’ and reverse primer 5’-CTCCTAGCGACCTAAC-3’. PCR products were purified and labeled using a BigDye Terminator 3.1 cycle-sequencing kit (Applied Biosystems, Foster City, CA), followed by sequencing in an ABI 3500XL Genetic Analyzer (Applied Biosystems).15 (link)
3
Quantification of Citrus Flavonoids
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They were as follows: Retsch MM400 Ball Mill(Laichi co., Germany); PTC200PCR(BIO-Rad, American); GelDox XR gel imaging system (BIO-Rad, American); DYY-8C Electrophoretic apparatus (Beijing Liuyi Instrument, China); BI3730XL sequenator (Applied Bio-systems, American); KRQ-300P manual climatic box (ChongQing YinBe Test Instrument, China); Plant DNA Extraction Kit (Tiangen Biotech Co., China); 2×Taq PCR MasterMix (Tiangen Biotech Co., China); primer was compounded by Sangon Co., China. Shimadzu; LC-20AT HPLC (Shimadzu Co., Japan); DM4000M microscope (Leica, Germany); Agilent 8453 ultraviolet and visible spectrophotometer (Agilent Technologies Co., American); hesperidin (MUST-12041206); narirutin (MUST-14081915); hesperetin (MUST-14100914); naringenin (MUST-14112310), four reference substance were bought from Chengdu Mansite biological technology co. LTD, purity>98.0%. The other reagents were analytically pure.
4
Detecting Frameshift Mutation in APC Gene
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Polymerase chain reaction (PCR) was performed to amplify the fragments of suspected candidate mutation loci, and Sanger DNA sequencing validation was performed to detect the corresponding loci in the proband and family members participating in the study. Primer Premier 5 software was used to design the target sequence primers. The APC sequence was obtained from GenBank (NM_000038.6), and the target amplicon length was 446 bp in APC exon 16 (c.3260_3261del: p.L1087fs). The following primers were used: F: 5′-TCAGATGAGCAGTTGAACTCTGGAAGG-3′ and R: 5′-CTATAATCAATAGGCTGATCCACATGAC-3′. The PCR was performed in a 50 µL reaction volume to amplify the target fragments on a thermocycler instrument (PTC-200 PCR, BioRad), and the annealing temperature of PCR was 58°C. The PCR products were purified using Takara reagents, and the PCR products of the target fragments were sequenced on an ABI 3730XL platform. The DNA extracted from the pedigree members (II1, II2, II8, II12, III1, III3, III4, III5, III6, III7, and III8) was subjected to PCR amplification and Sanger sequencing in the target region to detect whether they carried the frameshift mutation p.L1087fs.
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