opentrons OT-2 Workstations rau Cov Lus Qhia Ua Haujlwm Zoo Lab Automation

Npaj daim ntawv
Kev ntsuas thiab ntsuas ntsuas

Sau los ntawm Kennedy Bae, Ph.D. thiab Kinnari Watson, Ph.D.

Pib pib

Ua ntej yuav qhia zaj lus qhia, ua kom tiav cov kauj ruam hauv qab no ua ntej mus kawm.

Unbox lub OT-2
Teem lub Opentrons app
Txuas cov pipettes
Calibrate lub lawj
Calibrate taub hau ntev & pipette offset
Ntshuam txhua yam kev cai cuam tshuam rau lub app
Kuaj khiav cov txheej txheem ntawm OT-2

Xav tau kev pab ntxiv?

Rau kev txhawb nqa, thov mus saib peb Opentrons Help Center rau cov lus hais. Yog tias koj xav tau kev txhawb nqa ntxiv, thov hu rau support@opentrons.com.

Yog tias koj muaj lus nug ntsig txog txoj kev qhia, thov hu rau tus kws sau ntawv, Kennedy Bae, ntawm yog kennedy@opentrons.com, los yog Kinnari Watson, ntawm yog kinnari@opentrons.com.

Phau ntawv qhia
Kev ntsuas thiab ntsuas ntsuas

Sau los ntawm Kennedy Bae, Ph.D. thiab Kinnari Watson, Ph.D.

Lub hom phiaj

Lub chaw kuaj mob no yog tsim los tsim kom cov tub ntxhais kawm nkag siab txog kev siv cov txheej txheem dilutions los tsim cov qauv nkhaus, thiab kev siv cov qauv nkhaus rau kev sib cuam tshuam sample nqi. Qhov no suav nrog kev txaus siab ntawm:

• Basic pipeting txuj ci
• Serial dilutions
• Kev ntsuas cov ntawv luam tawm
• Ntau yam ntawm tus qauv nkhaus
• Kev siv cov txheeb cais los ntsuas qhov tseem ceeb

Cov tub ntxhais kawm ua phau ntawv pipetting nyob rau tib lub sijhawm nrog kev tuav cov kua dej, ua rau cov tub ntxhais kawm pom qhov ua tau zoo thiab muaj peev xwm ntawm ob txoj kev.

Cov Tub Kawm Ntawv

Lub chaw kuaj mob no tau tsim los siv rau hauv kev nkag mus rau cov chav kawm biology hauv nruab nrab. Nws yog qhov hloov pauv kom haum rau txhua tus tub ntxhais kawm uas tau cuv npe hauv chav kawm.

Keeb Kwm Paub

Cov tub ntxhais kawm yuav tsum muaj kev nkag siab txog kev nkag siab ntawm kev siv pipetting, spectrophotometric, qauv nkhaus, thiab kev txheeb xyuas cov ntaub ntawv.

Core Competencies

Laboratory Skills
Pipetting, sample kev npaj, thiab kev siv cov cuab yeej kuaj automation

Kev txheeb xyuas cov ntaub ntawv
Txhim kho cov qauv nkhaus, kev cuam tshuam, thiab kev txheeb xyuas kev txheeb xyuas

Kev xav zoo
Kev txhais cov ntaub ntawv sim, kev daws teeb meem, thiab kos cov lus xaus

Cov khoom siv

Opentrons raws tu qauv

Download tau raws tu qauv los ntawm
https://protocols.opentrons.com/protocol/customizable_s erial_dilution_ot2

• Xaiv cov parameters hauv qab no ntawm lub webnplooj ntawv raws li koj qhov teeb tsa thiab cov khoom siv:
  • Pipette Hom
  • Mount Sab
  • Hom tswv yim
  • Trough Hom
  • Hom phaj
• Khaws cov teeb tsa default rau:
  • Dilution factor (3)
  • Tus naj npawb ntawm dilution (10)
  • Tag nrho cov khoom ntim (150)
  • Pleev nyob rau hauv lub phaj zoo (yog)
  • Tswv yim siv tswv yim (siv ib lub tswv yim)
  • Volume ntawm huab cua sib txawv (10)

Cov khoom siv Opentrons

Opentrons OT-2 Automated Liquid Handling Robot
Opentrons p300 8-channel pipette

Cov Khoom Siv Tsis-Opentrons

Plate-based spectrophotometer
Software rau kev txheeb xyuas cov ntaub ntawv (ex. Excel lossis nplooj siab txheeb cais software)
P1000 sib txawv ntim phau ntawv pipettes (ib tus menyuam kawm ntawv)
P100 kuj tsis paub meej ntim phau ntawv pipette (tsuas yog ib qho yuav tsum tau ua, ua qauv qhia Part A)

Labware

Opentrons 300 μL Ntiv Rack
12-zoo trough
96-zoo Flat hauv qab phaj
Test tubes (6 rau ib tus menyuam kawm ntawv)
Dropper (1 rau tus menyuam kawm ntawv)

Reagents

1000 mL diH20 ib tus menyuam kawm ntawv
FD&C Blue No. 1 (McCormick® tau siv rau hauv kev txhim kho lub chaw kuaj ntshav no)

Kev sim Duration

Yuav tsum muaj Chav Kawm Sessions
1

Lab Run Sijhawm
Lub sijhawm kwv yees tag nrho: 2.5-3 teev
Taw qhia thiab Calibration Curve Generation: 35 feeb
Sample Kev Npaj thiab Pipetting: 1 teev
Cov ntaub ntawv khaws cia: 30 feeb

Kev daws teeb meem yooj yim

1. Ua ib qho kev sim ua ntej chav kawm; Txoj kev no yuav daws tau qhov xwm txheej tsis tau npaj tseg ua ntej cov tub ntxhais kawm tuaj txog.
2. Cov teeb meem nrog cov lus qhia tawm tsam daim hlau yuav luag txhua zaus vim yog siv lwm lub labware lossis cov neeg hlau calibration. Yog tias koj ntsib qhov no thiab tau lees paub qhov tseeb labware, sim rov ntsuas tus neeg hlau.
3. Yog tias koj xav tau hu rau Opentron Support, thov qhia rau lawv tias koj yog ib feem ntawm peb qhov kev pab cuam Opentrons rau Kev Kawm thiab hnub ntawm koj chav kuaj tom ntej.

Yuav tsum tau ua haujlwm ua ntej-lab

Ua ntej pib qhov kev sim no, cov tub ntxhais kawm yuav tsum muaj peev xwm thiab kev paub theoretical hauv qab no:

• Thawb lub pipette taub rau ntawm phau ntawv pipette
• Volume adjustments rau phau ntawv pipettes
• Kev paub txog kev paub txog spectrophotometer - Tus kws qhia ntawv yuav khiav lub spectrophotometer rau qhov chaw kuaj no
• Kev nkag siab ntawm cov qauv nkhaus, suav nrog kev paub txog qhov sib npaug uas txhais cov nkhaus, thiab muaj peev xwm los cuam tshuam cov txiaj ntsig
• Kev nkag siab zoo ntawm kev txheeb xyuas cov lej yooj yim, suav nrog suav thiab txhais cov ntsiab lus, tus qauv sib txawv, thiab R²

Cov txheej txheem qhia

1. Lab Introduction ~ 15 feeb

Qhov zoo tshaj plaws, cov tub ntxhais kawm yuav tsum nyeem phau ntawv qhia kev kuaj ua ntej tuaj rau hauv chav kawm, txawm li cas los xij, npaj siv sijhawm 10 feeb los tham txog cov txheej txheem pipetting, review ntawm standard curves, thiab interpolating sample nqi.

2. Calibration Curve Generation ~ 20 feeb

Tom ntej no, ua kom pom tus qauv nkhaus tsim (Tshooj A) rau cov chav kawm (lub chaw kuaj mob no xav tias tus kws qhia ntawv yuav yog tus tswv ntawm OT-2 neeg hlau thiab spectrophotometer):

1. Npaj cov tshuaj zas xim xiav nrog 50 μL ntawm cov zaub mov xiav xim thiab 10 mL ntawm diH2O.
2. Ntxiv 200 μL ntawm cov xim xiav dye tov rau lub qhov dej A1-H1 ntawm 96-zoo ca hauv qab phaj.
3. Ntxiv 20 mL dej rau A1 qhov dej ntawm lub trough reservoir.
   (Faj seeb: Cov kab tom qab qhov kawg dilution yog qhov chaw nyob ntawm qhov khoob. Qhov kawg ntawm lub qhov dej / lub pas dej yog cov dej khib nyiab.)
4. Npaj labware raws li hauv qab no:
   a. Qhov 1 = Opentrons 300 μL taub khib
   b. Qhov 2 = 12-channel reservoir
   c. Qhov 3 = 96-zoo tiaj tiaj hauv qab phaj
5. Nkag mus rau OT-2 khiav app los khiav cov txheej txheem.
6. Ntsuas qhov nqus ntawm 96-zoo phaj ntawm 450 nm.

3. Kev sib tham ~ 5 feeb

Tom qab khiav cov serial dilutions thiab tsim cov qauv nkhaus, koj tuaj yeem xav rov qab los ua ke hauv chav kawm thiab sib tham txog qhov muaj zog thiab lub sijhawm ntawm automation.

4. Ntu B: Phau Ntawv Sample Kev Npaj ~ 1 teev

Thaum lub sij hawm qhov chaw kuaj no, cov tub ntxhais kawm yuav raug hais kom nws tus kheej ua tiav Tshooj B ntawm qhov chaw sim:

1. Siv ib lub raj kuaj kom sau tau 200 μL aliquot ntawm cov xim muaj zog.
2. Siv lub raj thib ob kom sau 4 mL ntawm diluent (diH2O).
3. Pipette tsawg kawg yog 800 μL ntawm diH20 rau hauv txhua qhov ntawm 4 lub raj seem. Cov no yuav dhau los ua koj samples.
4. Siv lub dropper rau dispense ntau npaum li cas ntawm cov concentrated dye rau hauv txhua sampl cev.
5. Pipette 200 μL ntawm koj thawj sampnyob rau hauv lub qhov dej ntawm 96-zoo phaj. Rov ua qhov no 3 zaug ntxiv kom koj muaj 4 replicates ntawm qhov sample hauv 4 qhov dej ntawm lub phaj. Sau cov qhov dej uas koj tso no sample rau hauv (koj yuav zoo li yuav tau muab koj lub phaj 96-zoo nrog rau lwm tus menyuam kawm ntawv – koj xav paub seb qhov twg samples yog koj li!).
6. Rov ua cov kauj ruam saum toj no rau koj qhov seem 3 samples. Koj yuav tsum tau 4 qhov dej ntawm txhua qhov sample, rau tag nrho ntawm 16 lub qhov dej, tau ntim rau hauv lub phaj 96.
7. Ntsuas qhov nqus ntawm 96-zoo phaj ntawm 450 nm.

Thaum cov tub ntxhais kawm ua tiav Tshooj B, npaj rau kev ncig ncig lub chaw sim los teb cov lus nug thiab saib xyuas cov tub ntxhais kawm cov txheej txheem pipetting, ceeb toom cov tub ntxhais kawm kom sau cov qhov dej uas lawv siv, thiab muab kev pab rau tus kheej raws li xav tau.

5. Cov ntaub ntawv khaws cia ~ 30 feeb

Cia 30 feeb rau kev sau cov ntaub ntawv.

6. Kev tawg ~ 10 feeb

Tso lub sij hawm thaum kawg ntawm qhov chaw kuaj sim los piav qhia luv luv ntawm daim ntawv qhia chaw kuaj thiab muab sijhawm txaus rau cov tub ntxhais kawm los ntxuav lawv cov chaw nres tsheb thiab nug cov lus nug ntxiv.

Lab Report

Cov lus qhia

Muab cov tub ntxhais kawm npaj ib daim ntawv qhia txog kev kuaj sim uas suav nrog cov ntaub ntawv calibration curve, interpolated sample values, thiab statistical analysis. Muab cov txheej txheem rau kev tshaj tawm cov qauv thiab cov ntaub ntawv nthuav qhia. Qee lub tswv yim rau kev tshawb nrhiav hauv qab no.

• Tsim tus qauv nkhaus
  • Sau tus kheej tus nqi rau koj cov qauv.
  • Nrhiav qhov nruab nrab thiab tus qauv sib txawv hauv txhua qhov.
  • Siv cov nqi nruab nrab los txhais cov qauv nkhaus.
  • Txiav txim siab seb qhov kev ntsuas nkhaus haum rau cov ntaub ntawv cov ntsiab lus los ntawm kev suav tus nqi R2. Nco qab tias ib kab uas haum rau cov ntsiab lus cov ntaub ntawv zoo kawg nkaus muaj tus nqi R2 ntawm 1. Nws yog ib qho tseem ceeb los tsim cov qauv nkhaus tshiab nrog tus nqi R2 rau txhua qhov kev sim.
• Interpolation ntawm Sample Values
  • Siv koj tus qauv ntsuas nkhaus los cuam tshuam sample nqi manually.
  • Dab tsi yog qhov sib txawv ntawm tus qauv ntawm txhua tus ntawm koj cov samples?
  • Dab tsi yog qee qhov laj thawj rau qhov sib txawv no?
  • Dab tsi yog qhov txiaj ntsig ntawm kev ntsuas cov ntawv luam tawm?
• Dab tsi tiam sis koj tuaj yeem soj ntsuam ntawm cov sib txawv ntawm samples? Puas yog ib qho ntawm cov kev soj ntsuam sib txawv tseem ceeb?
  • Puas muaj cov kws tshaj lij? Yuav ua li cas siv cov txheeb cais tso cai rau kev ntsuam xyuas ntawm outliers vs cherry-xaiv cov ntaub ntawv?

Tub Kawm Ntawv Qhia
Kev ntsuas thiab ntsuas ntsuas

Lub hom phiaj

Lub chaw kuaj no yuav txhim kho koj txoj kev nkag siab ntawm kev siv serial dilutions los tsim cov qauv nkhaus, thiab kev siv cov qauv nkhaus rau kev sib cuam tshuam sample nqi. Qhov no suav nrog kev txaus siab ntawm:

• Basic pipeting txuj ci
• Serial dilutions
• Kev ntsuas cov ntawv luam tawm
• Ntau yam ntawm tus qauv nkhaus
• Kev siv cov txheeb cais los ntsuas qhov tseem ceeb

Hauv chav kuaj no cov tub ntxhais kawm yuav siv cov ntaub ntawv tau txais los ntawm kev siv cov raj xa dej nrog rau OT-2 automated kua handler, tso cai rau cov tub ntxhais kawm pom qhov ua tau zoo thiab muaj peev xwm ntawm ob txoj kev.

Cov Khoom Siv Yuav Tsum Tau

• Opentrons OT-2 Automated Liquid Handling Robot
• Opentrons p300 8-channel pipette
• Opentrons 300 μL Ntiv Rack
• Phau ntawv pipette (P1000)
• 12-zoo trough
• 96-zoo Flat hauv qab phaj
• 1000 mL deionized dej (diH2O)
• FD&C Blue No. 1 (McCormick® tau siv rau hauv kev txhim kho lub chaw kuaj ntshav no)
• 6 Test Tubes
• Dropper

Kev sim txheej txheem

Ntu A: Saib xyuas Standard Curve Generation

Lub OT-2 yuav ua automated pipetting ntawm serial dilutions. Thaum koj tus kws qhia ntawv ntsuas cov qauv no nrog lub spectrophotometer lawv yuav muaj peev xwm tsim tau tus qauv nkhaus.

Koj tus kws qhia ntawv yuav ua raws li cov lus qhia no los npaj thiab khiav OT-2 neeg hlau. Cov kauj ruam 1-4 yuav ua tiav manually (los ntawm koj tus kws qhia ntawv lossis cov tub ntxhais kawm tuaj yeem pab dawb) ua ntej ua haujlwm OT-2 cov kauj ruam neeg hlau:

1. Npaj cov tshuaj zas xim xiav nrog 50 μL ntawm cov zaub mov xim xiav thiab 10 mL ntawm diH2O.
2. Ntxiv 200 μL ntawm cov xim xiav dye tov rau lub qhov dej A1-H1 ntawm 96-zoo ca hauv qab phaj.
3. Ntxiv 20 mL dej rau A1 qhov dej ntawm lub trough reservoir.

(Faj seeb: Cov kab tom qab qhov kawg dilution yog qhov chaw nyob ntawm qhov khoob. Qhov kawg ntawm lub qhov dej / lub pas dej yog cov dej khib nyiab.)

4. Npaj labware raws li hauv qab no:

ο Qhov 1 = Opentrons 300 μL taub khib
ο Qhov 2 = 12-channel reservoir
ο Qhov 3 = 96-zoo hauv qab phaj

5. Nkag mus rau OT-2 khiav app khiav cov txheej txheem automated.

ο Download raws tu qauv los ntawm
https://protocols.opentrons.com/protocol/customiza ble_serial_dilution_ot2

• Xaiv cov parameters hauv qab no ntawm lub webnplooj ntawv raws li koj qhov teeb tsa thiab cov khoom siv:
  • Pipette Hom
  • Mount Sab
  • Hom tswv yim
  • Trough Hom
  • Hom phaj
• Khaws cov teeb tsa default rau:
  • Dilution factor (3)
  • Tus naj npawb ntawm dilution (10)
  • Tag nrho cov khoom ntim (150)
  • Pleev nyob rau hauv lub phaj zoo (yog)
  • Tswv yim siv tswv yim (siv ib lub tswv yim)
  • Volume ntawm huab cua sib txawv (10)

6. Ntsuas qhov nqus ntawm 96-zoo phaj ntawm 450 nm.

Note: Serial dilutions are performed by sequentially transferring aliquots of a standard into increasing amounts of diluent (see diagram). The OT-2 will aspirate an aliquot of the concentrated standards and dispense it into the diluent. It will then mix the concentrated dye and the diluent by repeatedly pipetting the liquid up and down. Then the robot will aspirate an aliquot of the first mixed standard and dispense it into the next column of diluent.

Ntu B: Kev coj Phau ntawv Sample Kev Npaj

1. Siv ib lub raj kuaj kom sau tau 200 μL aliquot ntawm cov xim muaj zog.
2. Siv lub raj thib ob kom sau 4 mL ntawm diluent (diH2O).
3. Pipette tsawg kawg yog 800 μL ntawm diH20 rau hauv txhua qhov ntawm 4 lub raj seem. Cov no yuav dhau los ua koj samples.
4. Tsim txawv samples nrog ntau qhov sib txawv ntawm cov concentrated piv rau diluent: Siv lub dropper los faib cov nqi sib txawv ntawm cov concentrated dye rau hauv txhua lub sampl cev.
5. Pipette 200 μL ntawm koj thawj sampnyob rau hauv lub qhov dej ntawm 96-zoo phaj. Rov ua qhov no 3 zaug ntxiv kom koj muaj 4 replicates ntawm qhov sample hauv 4 qhov dej ntawm lub phaj. Sau cov qhov dej uas koj tso no sample mus rau hauv (koj yuav zoo li yuav muab koj lub phaj 96-zoo nrog rau lwm cov tub ntxhais kawm koj xav paub qhov twg samples yog koj li!).
6. Rov ua cov kauj ruam 5 thiab 6 rau koj qhov seem 3 samples, yog li koj xaus nrog 4 replicates ntawm txhua sampua 4vl. Koj yuav tsum tau 4 qhov dej ntawm txhua qhov sample, rau tag nrho ntawm 16 lub qhov dej, tau ntim rau hauv lub phaj 96.
7. Ntsuas qhov nqus ntawm 96-zoo phaj ntawm 450 nm.

Tub Kawm Ntawv Quiz
Kev ntsuas thiab ntsuas ntsuas

Tub Kawm Ntawv Quiz

1. Dab tsi yog qhov txiaj ntsig ntawm kev siv serial dilutions los tsim cov qauv nkhaus?
2. Vim li cas cov tiam ntawm calibration curves tseem ceeb hauv kev ntsuas roj ntsha?
3. Sau npe ib qho kev sib tw hauv kev tsim cov calibration nkhaus thiab qhia txog kev daws teeb meem.
4. Piav qhia lub luag haujlwm ntawm cov cuab yeej siv hauv chav kuaj no.
5. Dab tsi yog qhov chaw ntawm kev hloov pauv los ntawm phau ntawv pipetting?
6. Vim li cas nws thiaj li muaj txiaj ntsig los ntsuas qhov rov ua dua ntawm txhua tus sample?
7. Dab tsi yog lub hom phiaj ntawm interpolating sample qhov tseem ceeb hauv qhov kev sim no?
8. Yuav ua li cas statistical tsom xam pab txhais qhov sib txawv ntawm ntau yam samples?
9. Yuav ua li cas cov kev tshawb pom los ntawm lub chaw kuaj ntshav no cuam tshuam rau kev tshawb fawb txog roj ntsha?

Cov ntaub ntawv / Cov ntaub ntawv

opentrons OT-2 Workstations rau Efficient Lab Automation [ua pdf] Cov lus qhia OT-2 Workstations rau Efficient Lab Automation, OT-2, Workstations rau Efficient Lab Automation, Efficient Lab Automation, Lab Automation, Automation

Cov ntaub ntawv

• Cov neeg siv phau ntawv