opentrons OT-2 Workstations don Ingantattun Umarnin Automation Lab

Prep Sheet
Ma'auni da Ƙididdigar Ƙididdiga

Kennedy Bae ne ya rubuta, Ph.D. da Kinnari Watson, Ph.D.

Farawa

Kafin koyar da shirin darasi, kammala waɗannan matakai kafin aji.

Cire akwatin OT-2
Saita aikace-aikacen Opentrons
Haɗa pipettes
Daidaita bene
Calibrate tsayin tip & biya diyya
Shigo da duk wasu ƙa'idodi masu alaƙa zuwa ƙa'idar
Gwada gudanar da yarjejeniya akan OT-2

Bukatar Ƙarin Taimako?

Don tallafin fasaha, da fatan za a duba mu Cibiyar Taimakon Opentrons don labarai masu dacewa. Idan kuna buƙatar ƙarin tallafi, tuntuɓi support@opentrons.com.

Idan kuna da tambayoyi masu alaƙa da shirin darasi, da fatan za ku tuntuɓi marubuta, Kennedy Bae, a kennedy@opentrons.com, ko Kinnari Watson, a kinnari@opentrons.com.

Jagoran Malamai
Ma'auni da Ƙididdigar Ƙididdiga

Kennedy Bae ne ya rubuta, Ph.D. da Kinnari Watson, Ph.D.

Manufar

Wannan dakin binciken yana da nufin haɓaka fahimtar ɗalibi na yin amfani da serial dilutions don samar da daidaitaccen lanƙwasa, da kuma amfani da madaidaitan lanƙwasa don haɗa s.ampda dabi'u. Wannan ya haɗa da godiya:

• Basic basirar bututu
• Serial dilutions
• Auna kwafi
• Kewayon daidaitattun lanƙwasa
• Amfani da kididdiga don tantance mahimmanci

Dalibai suna yin bututun hannu a layi daya tare da sarrafa ruwa ta atomatik, baiwa ɗalibai damar ganin ƙarfi da damar fasahohin biyu.

Masu Sauraron Dalibi

An tsara wannan dakin gwaje-gwaje don amfani da shi wajen shiga cikin manyan darussan nazarin halittu na farko. Yana da sauƙi don ɗaukar kowane adadin ɗaliban da aka yi rajista a cikin aji.

Ilimin Baya

Ya kamata ɗalibai su sami fahimtar ra'ayi game da pipetting, spectrophotometry, madaidaitan lanƙwasa, da bincike na ƙididdiga.

Ƙwararrun Ƙwararru

Ƙwararrun Ƙwararru
Bututu, sample shirye-shiryen, da kuma amfani da dakin gwaje-gwaje sarrafa kayan aiki

Binciken Bayanai
Haɓaka madaidaitan lanƙwasa, interpolation, da ƙididdigar ƙididdiga

Mahimman Tunani
Fassarar bayanan gwaji, magance matsala, da kuma zana madaidaicin ƙarshe

Kayayyaki

Opentrons Protocol

Zazzage yarjejeniya daga
https://protocols.opentrons.com/protocol/customizable_s erial_dilution_ot2

• Zaɓi sigogi masu zuwa akan webshafi bisa ga saitin ku da kayan aiki:
  • Nau'in Pipette
  • Dutsen Side
  • Nau'in Tukwici
  • Nau'in Tafarki
  • Nau'in Plate
• A kiyaye tsoffin saitunan don:
  • Matsalolin dilution (3)
  • Adadin dilution (10)
  • Jimlar ƙarar hadawa (150)
  • Blank a cikin farantin rijiya (ee)
  • Hanyar amfani da tukwici (amfani da tukwici ɗaya)
  • Girman tazarar iska (10)

Kayan Aikin Opentron

Robot mai sarrafa Liquid Mai sarrafa kansa na Opentrons OT-2
Opentrons p300 8-tashar pipette

Kayayyakin Mara-Mabude

spectrophotometer na tushen farantin
Software don nazarin bayanai (misali Excel ko software na ƙididdiga)
P1000 m girma manual pipettes (daya kowane dalibi)
P100 m girma na manual pipette (daya kawai ake buƙata, don nuna Sashe na A)

Labware

Opentrons 300 μL Tip Rack
12 - ruwa mai kyau
96-riji Flat Bottom farantin
Bututun gwaji (6 kowane ɗalibi)
Dropper (1 kowane dalibi)

Reagents

1000 ml diH20 kowane ɗalibi
FD&C Blue No. 1 (An yi amfani da McCormick® wajen haɓaka wannan lab)

Tsawon Gwaji

Da ake Bukata Zaman Aji
1

Lokacin Run Lab
Kiyasta jimlar lokacin: 2.5-3 hours
Gabatarwa da Ƙwararren Ƙwararren Ƙwararren Ƙwararru: Minti 35
Sample Shiri da bututu: 1 hour
Tarin Bayanai: Minti 30

Magance matsalar asali

1. Yi gwajin gwaji kafin aji; ta haka za a iya magance duk wani abin da ba a zata ba kafin dalibai su zo.
2. Batutuwa tare da tukwici masu ɗaukar faranti kusan koyaushe suna faruwa saboda amfani da madadin labware ko daidaitawar mutum-mutumi. Idan kun fuskanci wannan kuma kun tabbatar da madaidaicin labware, gwada sake daidaita robot ɗin.
3. Idan kuna buƙatar tuntuɓar Taimakon Opentrons, da fatan za a sanar da su cewa kuna cikin shirinmu na Opentrons don Ilimi da kwanan watan aji na lab ɗin ku na gaba.

Ayyukan Pre-Lab da ake buƙata

Kafin fara wannan lab, ɗalibai yakamata su sami ƙwarewar fasaha masu zuwa da ilimin ka'idar:

• Loda titin pipette akan pipette na hannu
• Gyaran ƙara don pipettes na hannu
• Ilimin ka'idar spectrophotometry -Malamin zai yi amfani da spectrophotometer don wannan lab.
• Fahimtar madaidaitan lanƙwasa, gami da sanin ma'auni wanda ke bayyana ma'anar lanƙwasa, da kuma ikon daidaita dabi'u.
• Ƙarfin fahimtar ƙididdigar ƙididdiga ta asali, gami da ƙididdigewa da ma'anar fassara, daidaitattun sabani, da R²

Jagorar Hanya

1. Gabatarwar Lab ~ Minti 15

Da kyau, ya kamata ɗalibai su karanta jagorar lab kafin su zo aji, duk da haka, su shirya ɗaukar mintuna 10 suna tattaunawa kan dabarun bututu, sakeview na madaidaitan lankwasa, da kuma interpolating sample dabi'u.

2. Ƙwararren Ƙwararren Ƙwararru ~ Minti 20

Na gaba, nuna daidaitattun tsararrun lankwasa (Sashe na A) don ajin (wannan ɗakin binciken yana ɗaukan cewa malami ne zai zama ma'aikaci ɗaya tilo na robot OT-2 da spectrophotometer):

1. Shirya maganin rini mai launin shuɗi tare da 50 μL na launin abinci mai launin shuɗi da 10 ml na diH2O.
2. Ƙara 200 μL na maganin rini mai shuɗi zuwa rijiyoyin A1-H1 na farantin ƙasa mai rijiyar 96.
3. Ƙara 20 ml na ruwa zuwa rijiyar A1 na tafki.
   (Lura: ginshiƙi bayan dilution na ƙarshe shine asalin wurin da ba a taɓa gani ba. Rijiyar ƙarshe na trough/tafki ita ce tsohuwar sharar ruwa.)
4. Shirya labware kamar haka:
   a. Ramin 1 = Opentrons 300 μL tip tip
   b. Ramin 2 = tafki mai lamba 12
   c. Ramin 3 = 96-rijiya lebur kasa farantin
5. Ci gaba zuwa aikace-aikacen gudu na OT-2 don gudanar da yarjejeniya.
6. Ƙididdige ɗaukar nauyin farantin rijiyar 96 a 450 nm.

3. Tattaunawa ~ Minti 5

Bayan gudanar da serial dilutions da samar da daidaitaccen lanƙwasa, kuna iya fatan dawowa tare a matsayin aji kuma ku tattauna ƙarfi da damar sarrafa kansa.

4. Kashi na B: Manual Sample Shiri ~ 1 hour

A lokacin wannan ɓangaren ɗakin binciken, za a nemi ɗalibai su kammala Sashe na B na ɗakin binciken da kansu:

1. Yi amfani da bututun gwaji guda ɗaya don tattara 200 μL aliquot na rini mai ƙarfi.
2. Yi amfani da bututun gwaji na biyu don tattara 4 ml na diluent (diH2O).
3. Pipette aƙalla 800 μL na diH20 cikin kowane ɗayan bututu 4 da suka rage. Waɗannan za su zama samples.
4. Yi amfani da ɗigon ruwa don ba da ɗimbin yawa na rini mai ƙarfi cikin kowane sampku tube.
5. Pipette 200 μL na farkon sample cikin rijiyar farantin rijiyar 96. Maimaita wannan sau 3 don samun sau 4 na wannan sample a cikin rijiyoyin 4 na farantin. Rubuta rijiyoyin da kuka sanya wannan sample shiga (watakila za ku raba farantin rijiyar 96 tare da sauran ɗalibai - kuna son sanin wane samples naku ne!).
6. Maimaita matakin da ke sama don ragowar s3 ɗin kuamples. Kuna da rijiyoyi 4 na kowane sample, don jimlar rijiyoyi 16, cike a cikin farantin rijiyar 96.
7. Ƙididdige ɗaukar nauyin farantin rijiyar 96 a 450 nm.

Yayin da ɗalibai ke kammala Sashe na B, yi shirin zagayawa a cikin lab don amsa tambayoyi da lura da fasahar bututun ɗalibi, tunatar da ɗalibai su yi rikodin rijiyoyin da suke amfani da su, da ba da taimako na mutum gwargwadon buƙata.

5. Tarin Bayanai ~ Minti 30

Bada minti 30 don tattara bayanai.

6. Ragewar ~ Minti 10

Bar lokaci a ƙarshen dakin gwaje-gwaje don gabatar da rahoton lab a taƙaice da samarwa ɗalibai isasshen lokaci don tsaftace tashoshin su da yin sauran tambayoyin.

Rahoton Lab

Umarni

Sanya ɗalibai don shirya cikakken rahoton lab wanda ya haɗa da bayanan daidaitawa, interpolated sample dabi'u, da kididdiga bincike. Samar da jagororin tsarin rahoto da gabatar da bayanai. Wasu ra'ayoyin don bincike suna ƙasa.

• Kafa madaidaicin lankwasa
  • Haɗa ƙimar daidaiku don ma'aunin ku.
  • Nemo ma'ana da daidaitaccen karkata a kowane.
  • Yi amfani da ma'aunin ƙima don ayyana madaidaicin lanƙwasa.
  • Ƙayyade yadda lanƙwan ƙididdiga ta dace da ma'aunin bayanai ta hanyar ƙididdige ƙimar R2. Ka tuna cewa layin da ya dace da wuraren bayanan daidai yana da ƙimar R2 na 1. Yana da mahimmanci don ƙirƙirar sabon madaidaicin lanƙwasa tare da ƙididdige ƙimar R2 don kowane gwaji.
• Interpolation na Sampda Dabi'u
  • Yi amfani da madaidaicin madaidaicin lanƙwan ku don haɗa sample ƙimar da hannu.
  • Menene ma'auni tsakanin kwafi na kowane s ɗin kuamples?
  • Wadanne dalilai ne zasu iya haifar da wannan bambancin?
  • Menene amfanin auna kwafi?
• Waɗanne halaye za ku iya lura da su a cikin nau'ikan s daban-dabanamples? Shin ɗaya daga cikin bambance-bambancen da aka lura da su yana da mahimmancin ƙididdiga?
  • Shin akwai masu fita? Ta yaya yin amfani da kididdiga zai ba da izinin kima na masu fita da bayanan ceri?

Jagoran ɗalibi
Ma'auni da Ƙididdigar Ƙididdiga

Manufar

Wannan ɗakin binciken zai haɓaka fahimtar ku ta amfani da siriyal dilutions don samar da daidaitaccen lanƙwasa, da kuma amfani da madaidaitan lanƙwasa don haɗa s.ampda dabi'u. Wannan ya haɗa da godiya:

• Basic basirar bututu
• Serial dilutions
• Auna kwafi
• Kewayon daidaitattun lanƙwasa
• Amfani da kididdiga don tantance mahimmanci

A cikin wannan dakin gwaje-gwaje ɗalibai za su yi amfani da bayanan da aka samu ta hanyar bututun hannu da kuma na'urar sarrafa ruwa ta OT-2 mai sarrafa kansa, baiwa ɗalibai damar ganin ƙarfi da damar fasahohin biyu.

Kayayyakin da ake buƙata

• Robot mai sarrafa Liquid Mai sarrafa kansa na Opentrons OT-2
• Opentrons p300 8-tashar pipette
• Opentrons 300 μL Tip Rack
• pipette na hannu (P1000)
• 12 - ruwa mai kyau
• 96-riji Flat Bottom farantin
• 1000 ml ruwa mai narkewa (diH2O)
• FD&C Blue No. 1 (An yi amfani da McCormick® wajen haɓaka wannan lab)
• 6 Gwajin Bututu
• Mai saukewa

Tsarin Gwaji

Sashe A: Kula Standard Curve Generation

OT-2 za ta yi aikin bututun mai sarrafa kansa na serial dilutions. Lokacin da malamin ku ya auna waɗannan ma'auni tare da spectrophotometer za su iya samar da daidaitaccen lanƙwasa.

Malamin ku zai bi waɗannan umarnin don shirya da gudanar da robot OT-2. Matakai na 1-4 za a kammala su da hannu (da malaminku ko ɗaliban sa-kai) kafin aiwatar da matakan robot na OT-2 mai sarrafa kansa:

1. Shirya maganin rini mai shuɗi tare da 50 μL na launin abinci mai launin shuɗi da 10 ml na diH2O.
2. Ƙara 200 μL na maganin rini mai shuɗi zuwa rijiyoyin A1-H1 na farantin ƙasa mai rijiyar 96.
3. Ƙara 20 ml na ruwa zuwa rijiyar A1 na tafki.

(Lura: ginshiƙi bayan dilution na ƙarshe shine asalin wurin da ba a taɓa gani ba. Rijiyar ƙarshe na trough/tafki ita ce tsohuwar sharar ruwa.)

4. Shirya labware kamar haka:

Ramin 1 = Opentrons 300 μL tip tara
Ramin 2 = tafki mai lamba 12
Ramin 3 = 96-rijiya lebur kasa farantin

5. Ci gaba zuwa aikace-aikacen gudu na OT-2 don gudanar da yarjejeniya ta atomatik.

Zazzage yarjejeniya daga
https://protocols.opentrons.com/protocol/customiza ble_serial_dilution_ot2

• Zaɓi sigogi masu zuwa akan webshafi bisa ga saitin ku da kayan aiki:
  • Nau'in Pipette
  • Dutsen Side
  • Nau'in Tukwici
  • Nau'in Tafarki
  • Nau'in Plate
• A kiyaye tsoffin saitunan don:
  • Matsalolin dilution (3)
  • Adadin dilution (10)
  • Jimlar ƙarar hadawa (150)
  • Blank a cikin farantin rijiya (ee)
  • Hanyar amfani da tukwici (amfani da tukwici ɗaya)
  • Girman tazarar iska (10)

6. Ƙididdige ɗaukar nauyin farantin rijiyar 96 a 450 nm.

Lura: Ana yin dilution na jeri ta hanyar canja wurin aliquots na daidaitattun jeri zuwa yawan adadin diluent (duba zane). OT-2 za ta yi amfani da ƙayyadaddun ƙayyadaddun ma'auni kuma za su ba da shi a cikin diluent. Daga nan za ta haxa rini da aka tattara da kuma narkar da su ta hanyar yin bututun ruwa akai-akai sama da ƙasa. Sa'an nan na'urar za ta nemi wani aliquot na farko gauraye daidaitattun kuma ya ba da shi a cikin shafi na gaba na diluent.

Sashe na B: Gudanarwa Manual Sample Shiri

1. Yi amfani da bututun gwaji guda ɗaya don tattara 200 μL aliquot na rini mai ƙarfi.
2. Yi amfani da bututun gwaji na biyu don tattara 4 ml na diluent (diH2O).
3. Pipette aƙalla 800 μL na diH20 cikin kowane ɗayan bututu 4 da suka rage. Waɗannan za su zama samples.
4. Ƙirƙirar sampLes tare da nau'i daban-daban na maida hankali da mai narkewa: Yi amfani da digo don rarraba yawan adadin rini a cikin kowane s.ampku tube.
5. Pipette 200 μL na farkon sample cikin rijiyar farantin rijiyar 96. Maimaita wannan sau 3 don samun sau 4 na wannan sample a cikin rijiyoyin 4 na farantin. Rubuta rijiyoyin da kuka sanya wannan sample cikin (wataƙila za ku raba farantin rijiyar 96 ɗinku tare da sauran ɗaliban da kuke son sanin wane samples naku ne!).
6. Maimaita matakai na 5 da 6 don ragowar s 3 na kuamples, don haka kun ƙare da kwafi 4 na kowane sample in 4 rijiyoyi. Kuna da rijiyoyi 4 na kowane sample, don jimlar rijiyoyi 16, cike a cikin farantin rijiyar 96.
7. Ƙididdige ɗaukar nauyin farantin rijiyar 96 a 450 nm.

Tambayoyi na ɗalibi
Ma'auni da Ƙididdigar Ƙididdiga

Tambayoyi na ɗalibi

1. Menene fa'idar yin amfani da serial dilutions don samar da daidaitaccen lankwasa?
2. Me yasa ƙarni na gyare-gyaren gyare-gyare yake da mahimmanci a ma'aunin halitta?
3. Sunan ƙalubalen ƙalubale ɗaya mai yuwuwa wajen samar da matakan daidaitawa da ba da shawarar mafita.
4. Bayyana rawar kayan aikin da aka yi amfani da su a cikin wannan dakin gwaje-gwaje.
5. Wadanne hanyoyin samun sauye-sauye na kowa daga bututun hannu?
6. Me yasa yake da amfani don auna kwafi na kowane sampku?
7. Menene manufar interpolating sampko darajar a cikin wannan gwaji?
8. Ta yaya bincike na kididdiga zai taimaka wajen fassara bambance-bambance tsakanin samples?
9. Ta yaya binciken daga wannan dakin binciken zai iya dacewa da binciken nazarin halittu?

Takardu / Albarkatu

opentrons OT-2 Worktations for Ingantacciyar Lab Automation [pdf] Umarni OT-2 Ayyuka don Ingantacciyar Lab Automation, OT-2, Wuraren Aiki don Ingancin Lab Automation, Ingantacciyar Lab Automation, Lab Automation

Magana

• Manual mai amfani