Ticks were crushed, individually for adults and in pools of five for nymphs, by shaking with a bead beater (mixer mill MM301, Qiagen, Hilden, Germany) as previously described [42 (link)]. DNA was extracted using the Nucleospin Tissue kit according to the manufacturer’s instructions (Macherey-Nagel, Duren, Germany). Adults and nymph pools were eluted in a final volume of 50 μL. DNA extracts were then stored at -20°C until use. DNA extraction efficiency was confirmed in all samples with polymerase chain reaction (PCR) amplification of the 16S rRNA mitochondrial gene using tick-specific primers TQ16S+1F (5′-CTGCTCAATGATTTTTTAAATTGCTGTGG-3′) and TQ16S-2R (5′-ACGCTGTTATCCCTAGAG-3′), as described [43 (link)].
Mm301
Manufactured by Qiagen
Sourced in Germany
The MM301 is a laboratory equipment designed for the efficient extraction and purification of nucleic acids, such as DNA and RNA, from various sample types. It utilizes a magnetic bead-based separation technology to isolate and purify the target molecules. The MM301 provides a reliable and automated solution for sample processing, ensuring consistent and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
Nucleospin tissue kit, by Macherey-Nagel (1 mentions)
Rq1 dnase 1, by Promega (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Qiagen (1 mentions)
Sds 2, by Thermo Fisher Scientific (1 mentions)
Improm 2 reverse transcriptase, by Promega (1 mentions)
Abi prism 7700 sequence detection system, by Thermo Fisher Scientific (1 mentions)
Hprt1, by Integrated DNA Technologies (1 mentions)
2 protocols using mm301
1
Tick DNA Extraction and Amplification
2
Total RNA Extraction and RT-qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from 10 to 30 mg of tissue or from treated cells using 1 ml of TRIzol Reagent (Thermo Fisher Scientific), according to the manufacturer's instructions. Tissues in TRIzol Reagent were pre-homogenised at 30 Hz for 3 min using the homogeniser MM301 (Qiagen). RNA for each sample was treated with RQ1-DNaseI (Promega) and reversed-transcribed using Improm II Reverse Transcriptase (Promega) and random hexamer primers. For quantitative RT-PCR, 2 µl of single-stranded complementary DNA was mixed with TaqMan Universal PCR Master (Thermo Fisher Scientific) and assay-ondemand gene-specific products, using HPRT1 as the normalising gene (Integrated DNA Technology), and analysed on a ABI PRISM 7700 Sequence Detection System (Life Technologies). The calculation of threshold cycle (C t ) values was performed using the SDS 2.2 software (Applied Biosystems), after automatically setting the baseline and the threshold. Data were analysed using the 2 ÀΔΔCt method, as previously described (24) .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!