The total DNA of the bacterial colony was isolated using the Bacteria Genomic DNA Extraction Kit (TaKaRa Minibeast Ver.3.0) and the sample quality was ensured using NanoDrop ND-1000 microspectrophotometer (NanoDrop Technologies, DE, USA). Whole-genome sequencing was performed on an Illumina HiSeq PE150 platform. A-tailed ligated paired-end adaptors with PCR ampli ed 350 bp inserts were used for library construction at Beijing Novogene Bioinformatics Technology Co., Ltd. From the Illumina PCR adapter reads, the lowquality reads were ltered as a quality control step by the sequencing company. All good quality paired reads were assembled using the SOAP denovo (http://soap.genomics.org.cn/soapdenovo.html) into several scaffolds (Li et al. 2010 (link)). The ltered reads were subjected to gap-closing. The whole-genome shotgun project was deposited at GenBank (accession PRJNA504495, S. maltophilia).
Nanodrop nd 1000 microspectrophotometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The NanoDrop ND-1000 microspectrophotometer is a compact and versatile laboratory instrument designed for the measurement of various samples. Its core function is to accurately measure the absorbance of small-volume liquid samples, enabling the quantification of DNA, RNA, and protein concentrations.
Automatically generated - may contain errors
Lab products found in correlation
Hiseq pe150, by Illumina (2 mentions)
Bacteria genomic dna extraction kit, by Takara Bio (2 mentions)
Sybr premix ex taq 2 kit, by Takara Bio (1 mentions)
Steponeplus, by Thermo Fisher Scientific (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
Minibest universal rna extraction kit, by Takara Bio (1 mentions)
3 protocols using nanodrop nd 1000 microspectrophotometer
1
Whole-Genome Sequencing of S. maltophilia
2
Whole-Genome Sequencing of S. maltophilia
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The total DNA of the bacterial colony was isolated using the Bacteria Genomic DNA Extraction Kit (TaKaRa Minibeast Ver.3.0) and the sample quality was ensured using NanoDrop ND-1000 microspectrophotometer (NanoDrop Technologies, DE, USA). Whole-genome sequencing was performed on an Illumina HiSeq PE150 platform. A-tailed ligated paired-end adaptors with PCR ampli ed 350 bp inserts were used for library construction at Beijing Novogene Bioinformatics Technology Co., Ltd. From the Illumina PCR adapter reads, the lowquality reads were ltered as a quality control step by the sequencing company. All good quality paired reads were assembled using the SOAP denovo (http://soap.genomics.org.cn/soapdenovo.html) into several scaffolds (Li et al. 2010 (link)). The ltered reads were subjected to gap-closing. The whole-genome shotgun project was deposited at GenBank (accession PRJNA504495, S. maltophilia).
3
Quantitative RT-PCR Analysis of Gene Expression
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Total RNA was extracted from 3T3-L1 cells using a MiniBEST universal RNA extraction kit (Takara Bio, Shiga, Japan). RNA concentrations were quantified using a NanoDrop ND-1000 Micro spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA), and reverse transcription was performed using 1 μg of total RNA and PrimeScript RT master mix (Takara) according to the manufacturer’s instructions. qRT-PCR was performed using a SYBR Premix Ex Taq II kit (Takara), with the amplification reaction monitored on a qPCR thermal cycler (Step One Plus; Applied Biosystems, Foster City, CA, USA). Relative expression was determined using the 2−ΔΔCT method, with levels normalized against those of 18S rRNA or the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase. Values are presented as fold changes compared to the control. The specific primers used in this study are listed in Table 1 .
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