Hielscher faka'ata tekinolosia

Ultrasonic Lysis ʻo Drosophila melanogaster sipinga

Drosophila melanogaster is widely used in laboratories as model organism. Therefore, pre-analytical preparation steps such as lysis, cell disruption, protein extraction and DNA shearing of Drosophila melanogaster samples must be frequently carried out. Ultrasonic dismembrators are reliable and efficient and can used to perform various tasks such as lysis, protein extraction or DNA fragmentation at ease by only adjusting ultrasonic process parameters. Ultrasonic homogenizers are thereby flexible tools with a broad range of applications.

Ultrasonic Lysis mo e ivi toʻo hingoa

Drosophila melanogaster is widely used as model organism in biological labs. Find here protocols for lysis, protein extracion and DNA shearing of D. melanogaster samples.Ko e Lysis, solubilization toʻotoʻo, homogenization mo e ivi toʻo ʻuli ko ha ngaahi ngaue angamaheni ia ki ultrasonic dismembrators ʻi he laboratories totonu. ʻOku feʻunga lelei ʻa e ultrasonic dismembrators mo e disruptors toʻotoʻo ki homogenize pepa holoholo ʻo e monumanu, ʻinisekite (e.g., Drosophila melanogaster, C. elegans) pe to ngaahi mataʻitohi. Ko hono fakaʻaongaʻi ko ia ʻo e ultrasonication ko e lysis ia ʻo e suspensions toʻotoʻo mo e pellets pea pehe ki hono toʻo ʻo e intracellular polotini.
Ko e ultrasonic lysis mo e ivi toʻo ʻuli ko ha founga falalaʻanga mo reproducible ia, ʻa ia ʻe lava ke fakahoko ʻo makatuʻunga ʻi he ngaahi founga kuo fokotuʻu. Koeʻuhi ʻe lava ke fakatonutonu ʻa e malohi ʻo e ultrasonic ʻo fakafou ʻi he ngaahi fakangatangata sonication hange ko e amplitude, siakale/sepo, mafana mo e sipinga ʻo e tohi, ʻe lava pe ke toutou fakahoko ʻaki ʻa e ola tatau.

Ngaahi lelei ʻo ultrasonic sipinga ʻo e teuteu

  • lelei 'aupito
  • Adjustable ki ha sipinga pau ʻo e naunau
  • Feʻunga mo ha faʻahinga tohi pe
  • Faitoʻo ʻoku ʻikai valaloto
  • ngaahi ola 'o e ngaahi
  • Faingofua mo malu

Ultrasonic DNA mo E RNA Fragmentation

After cell lysis and protein extraction, a common required step in sample preparation is the shearing and fragmentation of DNA, RNA, and chromatin, e.g. before chromatin immunoprecipitation (ChIP). DNA and RNA fragmentation can be reliably achieved by breaking the covalent bonds that hold the DNA together by physical forces. Using physical shearing such as sonication, at first the DNA strands are broken, then the DNA is fragmented into smaller pieces.
ʻOku falalaʻanga mo lelei ʻa e ultrasonic DNA fragmentation ʻi hono kamataʻi ʻo e DNA ki ha loloa pau, hange ko e 500bp (ngaahi hoa ʻi he fakavaʻe). ʻOku kau ʻi he ngaahi lelei lalahi ʻo ultrasonic DNA fragmentation ʻa hono puleʻi totonu ʻo e ngaahi fakangatangata mo e malohi ʻo e ultrasonic. ʻE lava ke fakatonutonu ʻa e ngaahi fakangatangata ʻo e ultrasonic ʻi hono fakatonutonu sonication malohi, siakale mo e taimi totonu. ʻOku malava heni ke faʻu ha tuʻunga kehekehe ʻo e DNA pea ʻe lava ke reliably faʻu mo hiki ʻa e loloa ʻo e DNA kuo fakataumuʻa ki ai. ʻOku toe lelei foki ultrasonic e fekauʻaki ʻa e DNA ke fakatupu ha kavenga mamafa molecular e DNA.

Ultrasonicator UP200Ht with microtip S26d2 for ultrasonic lysis of Drosophila samples

ultrasonicator UP200Ht with 2mm microtip S26d2 for the sonication of Drosophila samples

Kole 'a e fakamatala




Fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


Ultrasonic Lysis ʻo Drosophila melanogaster

Te ke lava ʻo maʻu ʻi lalo ha ngaahi ultrasonically kehekehe ki he lysis ʻoku tokoniʻi ʻe he ultrasonically, ivi toʻo ʻuli, mo e DNA pe chromatin fragmentation ʻo drosophila sipinga.

Ultrasonic Lysis ki hono fakafehokotaki ʻo e kolosi Immunoprecipitation (konga) lea

CLIP assay was performed as previously reported with some modifications. Approximately 20 mg ovaries from 0- to 1-day-old wild-type females were UV crosslinked (3 × 2000 μJ/cm2), homogenized on ice in 1 mL RCB buffer (50 mM HEPES pH 7.4, 200 mM NaCl, 2.5 mM MgCl2, 0.1% Triton X-100, 250 mM sucrose, 1 mM DTT, 1× EDTA-free Complete Protease Inhibitors, 1 mM PMSF) supplemented with 300 U RNAseOUT and placed on ice for 30 min. The homogenate was sonicated on ice, at 80% power, five times in 20 s bursts with a 60 s rest in between using the Hielscher Ultrasonic Processor UP100H (100 W, 30 kHz) mo centrifuged (16000 × g ʻi ha miniti ʻe 5 ʻi he 4 °C). Naʻe ʻosi maʻu ʻa e Soluble toʻo ʻaki ha μl e 20 ʻi he dynabeads ʻi ha miniti ʻe 20 ʻi he 4 °C. Hili hono toʻo e ngaahi sipinga ki he immunoblotting mo e quantitation ʻo e RNA input (1%), hp1 naʻe immunoprecipitated mo e kau ʻAnitai-HP1 9A9 antibody mei he 450 μl kuo ʻosi toʻo ʻe he incubation ki he 4 h mo e 50 μl ivi-Gdynabeads. Naʻe fufulu tuʻo 4 ʻa Immunoprecipitates mo RFB. Ke elute ʻa e immunoprecipitated RNAs, naʻe haka ʻa e pelleted ʻasi ʻi he 100 μL ʻo UltraPure DEPC-vai ʻi ha miniti ʻe 5. 900 μL Qiazol Reagent naʻe tanaki atu ia ki he supernatant teuteu ki he RNA. Naʻe fakaʻaongaʻi ʻa e RNA fakamaʻa ko ha template ke synthesize ʻa e FDNA ʻo fakaʻaongaʻi ʻa e oligo dT, hexamers mo superScript fulihi ʻa e III ʻo fakatatau ki he ngaahi founga ʻa e tokotaha ngaohi kapeti.
(Cassale et Al. 2019)

Ultrasonic Lysis for Chromatin Immunoprecipitation as

Chromatin immunoprecipitation was performed according to the method described by Menet with minor modifications. Approximately 20 mg ovaries from 0- to 1-day-old wild-type females were homogenized in 1 mL of NEB buffer (10 mM HEPES-Na at pH 8.0, 10 mM NaCl, 0.1 mM EGTA-Na at pH 8, 0.5 mM EDTA-Na at pH 8, 1 mM DTT, 0.5% NP-40, 0.5 mM Spermidine, 0.15 mM Spermine, 1× EDTA- free Complete Protease Inhibitors) with a homogenizer / immersion disperser 1 min (at 3000 rpm). The homogenate was transferred to a pre-chilled glass dounce and 15 full strokes were applied with a tight pestle. Free nuclei were then centrifuged at 6000xg for 10 min at 4°C. The nuclei-containing pellets were resuspended in 1 mL of NEB and centrifuged at 20000 × g for 20 min on sucrose gradient (0.65 mL of 1.6 M sucrose in NEB, 0.35 mL of 0.8 M sucrose in NEB). The pellet was resuspended in 1 mL of NEB and formaldehyde to a final concentration of 1%. Nuclei were cross-linked for 10 min at room temperature and quenched by adding 1/10 vol of 1.375 M glycine. The nuclei were collected by centrifugation at 6000 × g for 5 min. Nuclei were washed twice in 1 mL of NEB and resuspended in 1 mL of Lysis Buffer (15 mM HEPES-Na at pH 7.6, 140 mM NaCl, 0.5 mM EGTA, 1 mM EDTA at pH 8, 1% Triton X-100, 0.5 mM DTT, 0.1% Na Deoxycholate, 0.1% SDS, 0.5% N-lauroylsarcosine and 1× EDTA-free Complete Protease Inhibitors). Nuclei were sonicated using a Hielscher Ultrasonic Processor UP100H (100 W, 30 kHz) tuʻo ono ʻi he miniti ʻe 20 mo e miniti ʻe 1 ʻi he ʻaisi. Naʻe centrifuged ʻa Sonicated nuclei ʻi he 13000 × g ʻi ha miniti ʻe 4 ʻi he 4 °C. Ko e konga lahi ʻo e sonicated chromatin ko e 500 ki he 1000 (BP) hono loloa. ʻI he immunoprecipitation takitaha, naʻe incubated ai ha μg ʻe 15 ʻo chromatin ʻi he ʻao ʻo e 10 μg ʻo HP1 9A9 monoclonal antibody (3 h ʻi he 4 ° C ʻi ha veʻeteka vilo). ʻI he taimi ko ia, naʻ μl tanaki atu ʻa e dynabeads ʻe 50 ʻo e ivi ʻo e dynabeads G pea naʻe kei hokohoko atu pe incubation ʻi he po ko ia ʻi he 4 °C. Naʻe liʻaki ʻa e supernatants pea naʻe fufulu tuʻo ua ʻa e ngaahi sipinga ʻi Lysis ngataʻanga (fo takitaha miniti ʻe 15 ʻi he 4 ° C) pea tuʻo ua ʻi he TE ngataʻanga (1 mM EDTA, 10 mM TrisHCl ʻi he pH 8). Naʻe holoki ʻa Chromatin mei he ʻikai ke ʻi ai ha sitepu ʻe ua; ʻuluaki ʻi he 100 μl ʻo e eluition ngataʻanga 1 (10 mM EDTA, 1% SFU, 50 mM TrisHCl ʻi he pH 8) ʻi he 65 ° c ʻi ha miniti ʻe 15, hoko ai mo centrifugation mo fakaakeake ʻa e supernatant. Naʻe toe maʻu ʻa e naunau ne ʻasi ʻ μl 100 ʻo e TE + 0.67% S FUʻU. Naʻe incubated ʻa e eluate (200 μl) ʻi ha po kakato ʻi he 65 ° c ke fulihi ʻa e ngaahi fehokotakiʻanga ki he kolosi pea ngaohi ʻe he 50 μg/ml RNaseA ʻi ha miniti ʻe 15 ʻi he 65 ° c pea mo e 500 μg/ml Proteinase K ki he 3 h °C. Naʻe phenol ʻa e sipinga – chloroform maʻu mo ethanol meʻa--. Naʻe toe nofoʻi ʻa e DNA ʻi ha μl 25 ʻo e vai. Ke maximising ʻa e molecular pehe mo e DNA immunoprecipitated, naʻe toe lahi ange ʻa e kau kanititeiti afa ʻi ha optimized duplex-PCR Protocol ʻaki hano fakaʻaongaʻi ha ongo seti kehekehe ʻe ua ʻo e primers ʻi ha tali pe ʻe taha.
(Fakatau et Al. 2019)

UP200St TD_CupHorn he sonication ʻoku ʻikai fakahangatonu ʻo e ngaahi sipinga

UP200St TD_CupHorn ki he sonication ʻoku ʻikai fakahangatonu ʻo e ngaahi sipinga hange ko e DNA mo e chromatin fulufuluʻi

Ngaue ultrasonic toʻotoʻo Disruptors ki he ngaahi sipinga totonu

Ko Hielscher Ultrasonics ko ho hoa ngaue taimi loloa ia ʻi he taimi ʻoku hoko mai ai ki he ngaue maʻolunga ultrasonicators ki he fakahohaʻasi toʻotoʻo, lysis, ivi toʻo hingoa, DNA, RNA, mo e chromatin fragmentation pea pehe ki he ngaahi sitepu teuteu ki he teuteu ngaahi meʻa. ʻOku maʻu ʻe Hielscher ʻa e meʻangaue lelei taha ultrasonic ki hoʻo tohi kole mo e ngaahi fie maʻu totonu, ʻi hono fai ha lekooti fakaʻauliliki ʻo ultrasonic loki fakatotolo homogenizers mo e sipinga ʻo e ngaahi ʻiuniti teuteu.
Vakili-fa'ahinga insonifier UP200St ki he lysisTalafungani fekumi-taipeʻi ultrasonicator ʻaki ha kiʻi tokoni Micro hange ko e UP200St (200W; vakai ki he fakatata toʻohema) pe ko ha taha ʻo e sipinga ultrasonic sipinga teuteu ʻo e ngaahi ʻiuniti VialTweeter pe UP200ST_TD_CupHorn mo VialHolder ko ha kau faʻifaʻitakiʻanga favourite ʻi he fakatotolo mo e ngaahi meʻa laboratories. ʻOku lelei ʻa e ultrasonic ʻiloa fekumi, ʻi he taimi ʻoku teuteuʻi ai ha ngaahi sipinga siʻisiʻi ange kuo pau ke lysed, maʻu pe fragmented. ʻOku hanga ʻe he sipinga ʻo e ngaahi ʻiuniti teuteu VialTweeter mo UP200St_TD_CupHorn ʻo fakaʻata ʻa e hoko sonication ʻo aʻu ki he 10 pe 5.
Kapau ʻoku lahi ʻa e ngaahi fika (e.g. 96-ʻu lauʻi peleti vai, microtiter ʻu lauʻi peleti mo e ala meʻa pehee), kuo pau ke fai ha ngaue ki ai UIP400MTP Ko e fokotuʻutuʻu sonication lelei taha ia. ʻOku ngaue ʻa e UIP400MTP ʻo hange ha cuphorn lahi ange, ʻa ia ʻoku fonu ʻi he vai pea ʻoku ʻi ai ha feituʻu feʻunga ke maʻu ai ha ʻu lauʻi peleti Micro. ʻI hono fakalele ʻe ha 400 Watts malohi ultrasonic processor, ʻoku fakahoko ʻe he UIP400MTP ha teunga mo ha sonication lahi ʻo e ʻu lauʻi peleti fakavahaʻa-taʻu kae lava ke veuki e ngaahi selo, lyse sipinga, solubilize pellets, toʻo polotini pe kosi DNA.

Mapuleʻi totonu ʻo fakafou ʻi he polokalama Smart Software

Processors ngaue 'a e Hielscher 'o e ngaahi hdT 'e lava ke ongo'i fiemalie mo faka'onga'i fakalele 'o fakafou 'i he browser remote control.ʻOku fakanaunau ʻa e ngaahi founga Hielscher sonication kotoa pe mei he 200 Watts ʻaki ha naunau fakaʻilekitulonika colour ki he monitoa mo ha polokalama fakakomipiuta fakapotopoto. ʻOku fakahaofi he taimi pe ko ia ʻa e ngaahi fakangatangata ʻo e founga ngaue protocolling ultrasonic kotoa pe ʻi he faile CSV ʻi ha kaati SD ʻi he kamata pe ʻa e ultrasonicator. ʻOku hanga ʻe he meʻa ni ʻo ʻai ke toe faingofua ange ʻa e fakatotolo mo e protocolling. Hili e ngaahi ʻahiʻahi sonication pe sipinga teuteu, te ke lava pe ʻo toe vakaiʻi ʻa e ngaahi fakangatangata sonication ʻo e sonication takitaha pea fakafehoanaki kinautolu.
Via the intuitive menu, numerous parameter can be preset before sonication: For instance, to control the temperature in the sample and to prevent its thermal degradation, an upper limit of sample temperature can be set. A pluggable temperature sensor, which comes with the ultrasonic unit, gives the ultrasonic processor feedback on the actual sonication temperature. When the upper temperature limit is reached, the ultrasonic device pauses until the lower limit of the set ∆T is reached and starts then automatically sonicating again.
Kapau ʻoku fie maʻu ha sonication mo ha ivi pau, te ke lava ʻo pisitoni ʻa e ivi fakaʻosi ultrasonic ʻo e lele sonication. Ko e moʻoni, ʻe lava ke fokotuʻu fakafoʻituitui pe ʻa ultrasonic pulsation mo e cycle Mode.
Ke toe fakaʻaongaʻi hoʻo ngaahi fakangatangata sonication lelei taha, te ke lava ʻo seivi ʻa e ngaahi founga sonication kehekehe (e.g. sonication taimi, malohi, siakale ʻo e mode mo e ala meʻa pehe) ko ha founga kimuʻa, koeʻuhi ke lava ʻo faingofua pea toe kamata vave.
Ke maʻu ha faingamalie lahi ange, ʻe lava ke fakalele ʻa e ngaahi ʻiuniti ultrasonic kotoa pe ʻo fakafou ʻi he browser ʻoku ʻikai lava ke mapuleʻi ʻi ha faʻahinga browser angamaheni (e.g., InternetExplorer, Safari, Chrome etc.). Ko e fehokotakiʻanga LAN ko ha kiʻi fokotuʻutuʻu faingofua pe ia ʻo e plug-n pea ʻoku ʻikai fie maʻu ke toe fola ha polokalama fakakomipiuta kehe.
ʻOku tau ʻilo ʻi Hielscher ʻoku fie maʻu ʻe he sonication lelei ʻo e ngaahi sipinga totonu ʻa e totonu mo e repeatability. Ko ia ai, naʻa mau fokotuʻutuʻu ʻemau ultrasonicators ko ha ngaahi meʻangaue fakapotopoto mo e ngaahi meʻa kotoa pe ʻe lava ke maʻu ai ha teuteu lelei, falalaʻanga, reproducible mo faingamalie.

Fetuʻutaki mai he taimi ni ʻo fakamatala mai fekauʻaki mo hoʻo ngaahi sipinga totonu mo e ngaahi sitepu ʻoku fie maʻu ki he teuteu. Te mau fokotuʻu atu ʻa e sipinga ultrasonic feʻunga taha ʻo e meʻangaue teuteu mo tokoniʻi koe ʻi ha toe ngaahi fakamatala hange ko e ngaahi fokotuʻu mo e ngaahi fokotuʻu.

The table below gives you an indication of the approximate processing capacity of our ultrasonic systems from ultrasonic micro-tips and classic ultrasonic homogenizers to MultiSample ultrasonicators for the convenient, reliable preparation of numerous samples:

Kulupu (batch) 'o e tohi 'Oku tafe mai 'a e 'ea 'Oku fokotu'u atu 'a e ngaahi me'angaue
96-ngaahi lauʻi peleti/microtiter n.a. UIP400MTP
10 à 0.5 ki he 1.5 mL n.a. VialTweeter 'i he UP200St
CupHorn ki he sonication ʻoku ʻikai fakahangatonu, hange ko e 5 n.a. UP200ST_TD_CupHorn
0.01 ki he 250mL 5 ki he 100mL/miniti UP50H
0.01 ki he 500mL 10 ki he 200mL/miniti 'e UP100H
0.02 ki he 1L 20 ki he 400mL/miniti 'e UP200Ht / UP200St
mL 'i he 10 ki he 2000 20 ki he 400mL/miniti 'e UP200Ht, UP400St
0.25 ki 5L 0.05 ki he 1L/miniti UIP500hdT

Fetu'utaki mai kiate kimautolu! / Kole kiate kitautolu!

Kole ki ha fakamatala lahi ange

Kataki 'o ngaue 'aki e foomu 'i lalo ke kole ha toe fakamatala ki he ultrasonic processors, faka'aonga'i mo e totongi. Te mau fiefia ke alea'i e founga 'oku mo 'i ai mo ho'omou ngaahi fie ma'u ultrasonic he polokalama!









Kataki 'o fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


The VialTweeter is a MultiSample Ultraonicator that allows for reliable sample preparation under precisely controlled temperature conditions.

Ko e ʻiuniti teuteu ultrasonic fakavahaʻa-taʻu VialTweeter fakaʻata ʻa e hoko sonication ʻo aʻu ki ha foʻi vaio ʻe 10. ʻI he clamp ʻa e meʻangaue VialPress, ʻe lava ke lomiʻi ai ha toe tiupi ʻe 4 ki muʻa ʻi ha sonication lahi.

Ngaahi tohi/fakamo'oni fakafolofola



Hielscher Ultrasonics supplies high-performance ultrasonic homogenizers from lab to industrial size.

Ngaue ultrasonics maʻolunga! ʻOku ʻufiʻufi ʻe he koloa ʻa Hielscher ʻa e tuʻunga kakato mei he fale fakatotolo fakakemi ultrasonicator ʻi he ngaahi ʻiuniti ʻi ʻolunga ki he ngaahi polokalama ultrasonic lalahi.


Ngaahi mo'oni'i me'a 'oku mahu'inga ke 'ilo'i

Metabolomics

Metabolomics is the study of small molecules, known as metabolites, present within cells, biofluids, tissues or organisms. These small molecules and their interactions within a biological system are summarized under the umbrella term “metabolome” and the research field is called metabolomics. The metabolome research is closely connected with the rapidly emerging field of precision medicine. The understanding of the metabolome and its relation to various diseases helps to develop disease prevention and clinical care strategies whilst individual variability in environment, lifestyle, genetics, and molecular phenotype. In order to release metabolite molecules from cells, ultrasonication is frequently used in biological laboratories for pre-analytical sample preparation such as cell disruption, lysis and the extraction of proteins, lipids and other molecules.