Micro kit

Manufactured by Qiagen

Sourced in Germany, United Kingdom, United States

The Micro Kit is a compact and efficient laboratory equipment designed for the extraction and purification of nucleic acids, including DNA and RNA, from a variety of sample types. It utilizes a silica-based membrane technology to capture and purify the target molecules, providing a reliable and consistent method for sample preparation.

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Lab products found in correlation

89 protocols using micro kit

DNA Extraction from Frozen and Paraffin-Embedded Tissues

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For the cut-off study, DNA was extracted from frozen tissues at the Mayo Clinic Biospecimens Accession and Processing laboratory (Rochester, MN) using the Qiagen micro kit (Qiagen, Valencia, CA) prior to quantification by a Nanodrop spectrophotometer (Thermo Scientific, Wilmington, DE). Upon receipt of tissue-extracted DNA specimens in the research laboratory, quantity was verified by PicoGreen method (Molecular Probes, Eugene, OR). For the cohort study, DNA was extracted from paraffin-embedded tissues by laboratory personnel using the Qiagen micro kit, eluted into 100 µl of buffer and also quantified by the PicoGreen method.

Johnson D.H., Taylor W.R., Aboelsoud M.M., Foote P.H., Yab T.C., Cao X., Smyrk T.C., Loftus EV J.r., Mahoney D.W., Ahlquist D.A, & Kisiel J.B. (2016). DNA Methylation and Mutation of Small Colonic Neoplasms in Ulcerative Colitis and Crohn’s Colitis: Implications for Surveillance. Inflammatory bowel diseases, 22(7), 1559-1567.

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RNA-seq Analysis of Prostate Cancer Tissues

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For RNA-seq analyses, bulk prostate tissues, sorted LIN^-EpCAM⁺ prostate cancer cells and cancer- or spleen-associated CD45⁺CD3⁺CD8⁺ T cells were used. RNA was extracted by using RNAeasy mini kit (Qiagen 74106) or micro kit (Qiagen 74004), and cell line or tumor tissue cDNA library was constructed by using NEBNext® Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (E7530). The cDNA library of CD8⁺ T cell and cancer cell was constructed by using SMART-SEQ 2 protocol. Sequencing was performed by Illumina - HiSeq PE150.
All RNA-seq data were aligned to the mm10 genome using Tophat (version v2.1.1). Differentially expressed genes were identified by Cuffdiff (version v2.2.1)^{64 (link)}. FPKM was used for following analysis and comparison. GSEA analysis was performed as software suggested^{65 (link)}. T cell clonotype diversity analysis was performed by TRUST^{43 (link)}. The pathway activity score was calculated with GSVA⁶⁶. The expression levels of TLS score genes^{46 (link)} were scaled to the range of 0 to 1, and TLS score was defined as the mean scaled value of related genes for each individual sample,

Qi Z., Xu Z., Zhang L., Zou Y., Li J., Yan W., Li C., Liu N, & Wu H. (2022). Overcoming resistance to immune checkpoint therapy in PTEN-null prostate cancer by intermittent anti-PI3Kα/β/δ treatment. Nature Communications, 13, 182.

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Quantitative RT-PCR Gene Expression Analysis

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RNA was extracted with the QIAGEN RNeasy Plus Mini kit (cat# 74136) or Micro Kit (cat# 74034). cDNA was synthesized from RNA with the SuperScript III kit (Invitrogen, cat# 18080093). Transcripts were amplified with Power SYBR Green PCR Master Mix (Applied Biosystems, cat# 4367659), and quantitative PCR was performed on an Applied Biosystem ViiA 7 real-time PCR System. Relative expression level was calculated by the 2^^−(ΔΔCt) method in comparison to EEF1A1 control. Statistics was calculated using a Student’s t test against the control condition. Primers were designed using Primer-BLAST software.

Petrovic J., Zhou Y., Fasolino M., Goldman N., Schwartz G.W., Mumbach M.R., Nguyen S.C., Rome K.S., Sela Y., Zapataro Z., Blacklow S.C., Kruhlak M.J., Shi J., Aster J.C., Joyce E.F., Little S.C., Vahedi G., Pear W.S, & Faryabi R.B. (2019). Oncogenic Notch promotes long-range regulatory interactions within hyperconnected 3D cliques. Molecular cell, 73(6), 1174-1190.e12.

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Quantifying Mart1-specific CD8+ T Cells

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After TAME, 1500 naïve and memory Mart1-specific CD8⁺ T cells were sorted into RLT Buffer (Qiagen, France). Total RNA was extracted (Qiagen Microkit). cDNA were generated using the Supercript II enzyme (Invitrogen, France). RT-PCR reactions, thermal cycling conditions, calculations for relative usage of each Vβ family, and immunoscope profiles were performed as previously described (Alanio et al., 2013 (link); Bouvier et al., 2011 (link)) (Supplementary file 2).

Alanio C., Nicoli F., Sultanik P., Flecken T., Perot B., Duffy D., Bianchi E., Lim A., Clave E., van Buuren M.M., Schnuriger A., Johnsson K., Boussier J., Garbarg-Chenon A., Bousquet L., Mottez E., Schumacher T.N., Toubert A., Appay V., Heshmati F., Thimme R., Pol S., Mallet V, & Albert M.L. (2015). Bystander hyperactivation of preimmune CD8+ T cells in chronic HCV patients. eLife, 4, e07916.

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Quantitative Gene Expression Analysis

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Cells were lysed and homogenized using QIAGEN RLT buffer and Shredder columns before mRNA was extracted with the RNeasy Mini or Micro Kit (all QIAGEN). The cDNA was synthetized using the SuperScript VILO cDNA Synthesis kit (ThermoFisher), and gene expression was analyzed with the StepONE Plus PCR System by using pre-manufactured TaqMan primer together with the TaqMan Gene Expression Master Mix (all Applied Biosystems). The following TaqMan primer specificities were used (mouse Ltb: Mm00434774_g1, mouse Actb: Mm00607939_s1, human LTB: Hs00242739_m1, human B2M: Hs00187842_m1). Data were normalized to the housekeeping genes Actb or B2M.

Menzel L., Zschummel M., Crowley T., Franke V., Grau M., Ulbricht C., Hauser A., Siffrin V., Bajénoff M., Acton S.E., Akalin A., Lenz G., Willimsky G., Höpken U.E, & Rehm A. (2021). Lymphocyte access to lymphoma is impaired by high endothelial venule regression. Cell Reports, 37(4), 109878.

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Genomic DNA Extraction and PCR Amplification

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Genomic DNA was extracted by Micro Kit (QIAGEN) according to the manufacturer’s instructions. TaKaRa Taq^TM (Takara, R001A) was used for PCR amplification. Primers used in PCR were FR3-JH (FR3; 5’-CCGAGGACACGGCCGTGTATTACTG-3’ and JH; 5’-AACTGCTGAGGAGACGGTGACC-3’). PCR settings were designed based on the previous studies^{18 (link),59 (link)}. Images were obtained by iBright FL1000 (Thermo Fisher Scientific).

Harada T., Kikushige Y., Miyamoto T., Uno K., Niiro H., Kawakami A., Koga T., Akashi K, & Yoshizaki K. (2023). Peripheral helper-T-cell-derived CXCL13 is a crucial pathogenic factor in idiopathic multicentric Castleman disease. Nature Communications, 14, 6959.

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Transcriptome Analysis of Organoids

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Total RNA was extracted from organoids by RNeasy Mini Kit or Micro Kit (Qiagen) and cDNA generated with PrimeScript RT reagent Kit (Perfect Real Time) (Takara) according to the manufacturer’s instructions. qPCR was performed with a minimum of three biological replicates per gene using THUNDERBIRD SYBR qPCR Mix (Toyobo) according to the manufacturer’s instructions and ran on Mx3000P Real-Time QPCR System (Agilent). Analysis was carried out using double CT method. Primers used this study are listed in SI Appendix, Table S1.

Murakami K., Terakado Y., Saito K., Jomen Y., Takeda H., Oshima M, & Barker N. (2021). A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells. Proceedings of the National Academy of Sciences of the United States of America, 118(4), e2016806118.

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Quantifying Gene Expression in RPE Cells

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Total RNA was extracted from RPE cells using the RNEasy Mini or Micro Kit (Qiagen) with on-column DNase digestion. cDNA was synthesized using the High Capacity cDNA Synthesis kit (Applied Biosystems). Individual gene expression was assessed using predesigned Taqman assays (Applied Biosystems) and the reactions were carried out on the CFX96 iCycler platform (Biorad). Gene expression in all instances was quantified by the 2^-ΔΔCt relative quantification method [31 (link)] and normalized to geometric means of at least two housekeeping genes.

Choudhary P., Dodsworth B.T., Sidders B., Gutteridge A., Michaelides C., Duckworth J.K., Whiting P.J, & Benn C.L. (2015). A FOXM1 Dependent Mesenchymal-Epithelial Transition in Retinal Pigment Epithelium Cells. PLoS ONE, 10(6), e0130379.

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Dmxl2 Expression in Gonad Development

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We studied Dmxl2 expression during gonad development, by extracting total RNA from pools of ovaries or testes at different developmental stages, with the RNeasy Mini or Micro kit (Qiagen), depending on the amount of tissue. Three biological replicates were prepared for each stage and sex. The Maxima First-Strand cDNA Synthesis Kit (Thermo Scientific) was used to synthesize cDNA for RT-qPCR from 200 ng of RNA. RT-qPCR was performed in triplicate for all genes with the Absolute Blue SYBR Green ROX mix (Thermo Scientific), in the StepOnePlus Real-Time PCR System (Applied Biosystems). Based on the output of the GeNorm program, we used ActB, and Ywhaz as the reference genes for this study (S1 Table). The results were analyzed with qBase Software [61 (link)].

Gobé C., Elzaiat M., Meunier N., André M., Sellem E., Congar P., Jouneau L., Allais-Bonnet A., Naciri I., Passet B., Pailhoux E, & Pannetier M. (2019). Dual role of DMXL2 in olfactory information transmission and the first wave of spermatogenesis. PLoS Genetics, 15(2), e1007909.

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Quantitative Gene Expression Analysis

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Total RNA was extracted using either RNeasy Mini or Micro Kit together with the Qiashredder Kit (Qiagen, Hilden, Germany). An on-column DNAse treatment was included during the extraction. The following complementary DNA synthesis was performed using MultiScribe Reverse Transcriptase enzyme (Applied Biosystems, Foster City, CA) and random primers. Quantitative PCR was carried out in a 7300 Real-Time PCR (RT-PCR) System (Applied Biosystems) with SYBR Green PCR Master mix (Applied Biosystems). Comparative Ct method was used in order to quantify mRNA levels and three reference genes (SDHA, UBC, YWHAZ) were included to normalise gene-expression levels. Experiments were performed in triplicates. Primers are listed in Supporting information Table 2.

Braekeveldt N., Wigerup C., Gisselsson D., Mohlin S., Merselius M., Beckman S., Jonson T., Börjesson A., Backman T., Tadeo I., Berbegall A.P., Öra I., Navarro S., Noguera R., Påhlman S, & Bexell D. (2014). Neuroblastoma patient-derived orthotopic xenografts retain metastatic patterns and geno- and phenotypes of patient tumours. International Journal of Cancer. Journal International du Cancer, 136(5), E252-E261.

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