A Gilman P20 Pipet Set to a Reading of 152 Will Deliver

ane.3: Micropipetting

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36745

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• Source: ASCCC Open Educational Resources Initiative

Learning Objectives

Goals:

• Utilize various instruments plant in the biotechnology lab.
• Measure volume with precision and accuracy.
• Pipet with precision and accuracy.
• Learn how to utilise a micropipette to measure out very small volumes.

Student Learning Outcomes:

Upon completion of this lab, students volition be able to:

• Make accurate and precise measurements with micropipettes and serological pipettes.
• Summate percent error for a given measurement.
• Read, ready, and operate a micropipette.
• Make up one's mind which pipette should be used to measure a specific book.
• Decide how accurately you can measure with each micropipette.

Introduction to Micropipetting:

The ability to measure very small amounts, microliters (µl), of liquid chemicals or reagents is a fundamental skill needed in the biotechnology or enquiry lab. Scientists apply a device called a micropipette to measure out these very minor volumes with accuracy. This activity introduces the technique of micropipetting. Call up, as with all fine motor skills, this new skill will crave practice and determination. Be certain to operate the micropipette slowly and carefully.

Role I: Choosing and Setting the Micropipette

Effigy 1. Labeled parts of a micropipette, front and dorsum.

At that place are several sizes of micropipettes used in the biotechnology lab. Today, yous will exist using the P-1000, P-200, and P-20. The P-one thousand measures volumes between 100-thousand µl, the P-200 measures volumes between 20-200 µl, and the P-twenty measures volumes in the two-20 µl range. It is important to always option the correct micropipette for the volume to be measured.

Looking at Figure 3.1, you can see that each micropipette has a similar but unlike brandish window. For the P1000, the blood-red number indicates the thousands place, followed by the hundreds, tens, and the ones displayed as pocket-sized vertical lines. Each line represents 2 µl. The P-200 is reads differently. The display from the top downwardly reads, hundreds, tens, ones, and the vertical lines are considered 0.2 µl. Finally, the P-20 can be read from the acme downwards tens, ones, and the carmine tenths.

Effigy 2. Reading a micropipette at various sizes.

A. Choosing your Micropipette

For each amount listed below, bespeak the correct micropipette needed to measure the book accurately and then set the pipette to the indicated corporeality and show your partner.

Table 1. Pipette Size Choice

Amount	Pipette Needed		Partner Observation
567 µl:
ii. 160 µl:
3. 700 µl:
4. 25 µl:
v. fifteen µl:

B. Setting your Micropipette

Materials

• P-20 micropipette
• P-20 tips
• Waste container
• Tube of ruby-red dye in tube rack
• Laminated sheet for pipetting

Procedure

1. Each student will load 5, 10, 15, and 20 µl of carmine dye onto the laminated sheet.
2. Locate the p-20 and set the dial to 5 µl.
3. Concur the micropipette in your dominant manus, and gently but securely place the end of the micropipette into the proper size tip. Once the tip is on, be conscientious not to impact the tip on anything! If your tip touches the bench, lab coat etc. eject the tip into the waste container and become a new clean pipet tip.
4. With your other paw, open the cap of the tube of red dye and bring the tube of red dye to eye level,
5. Push the micropipette plunger down to the outset end and agree your thumb in this position.
6. Place the pipet tip into the ruddy dye solution.
7. Gently release your thumb from the plunger to draw fluid into the tip.
8. Confirm that the tip has liquid and that no bubbles are present within the tip.
9. Close the tube of red dye and place back in tube rack.
10. Gently touch the tip to the centre of the circumvolve labeled 5 µL and slowly push button all the way down (to second end) on the plunger to dispense the liquid.
11. Repeat this process for the remaining volumes.
12. Exist sure to watch your groupmates to provide feedback and aid with their technique.

Results

Have a picture or draw a picture of your spots and include this in your lab notebook every bit Figure 1. Brand sure the effigy has a title.

Determination

1. Observe if your spots were similar in size to your groupmates.
2. Which book had the most variability?
3. What could have contributed to your spot being too large or pocket-size?

Function 2: Pipetting Practice

A. Microplate Fine art

Materials

• p20 pipette (one)
• p200 micropipette (1)
• P-20/P-200 tips
• Microplate fine art set (design cards, colored dyes, and 96-well microplate) (one)
• Analytical or electronic balance

Process

1. Obtain a 96-well microplate, a design carte, and tubes of colored dyes.
2. Write the Microplate Art Design number in your lab notebook.
3. Using the gram residue, obtain the weight of your 96 well microplate and tape in your notebook.
4. Using the p200 micropipette with tip, dispense 50 µl of dye into the wells written on the design carte du jour.
5. Once you take finished pipetting, counterbalance your completed microplate, and record in your lab notebook.

Results

1. Be sure to record your weight in grams of your microplate pre/post pipetting in your lab notebook.
2. Using these values, calculate your percent error of the microplate yous but created. Include the calculation in your lab notebook.
3. Accept a moving-picture show of your microplate blueprint and include this in your lab notebook.

CONCLUSION

1. Was your percentage error below +/- 5%? If your percent fault was above this range, elaborate on the potential causes.
2. Did your pattern expect right? How could you avoid errors in the future?

B. Micropipette Practice Matrix

Materials

• p20 pipette (1)
• p200 micropipette (1)
• 1.5ml microfuge tubes (3)
• Permanent marker
• Analytical or electronic balance

Procedure

1. Label 3 microfuge tubes: 1, 2, 3,
2. Weigh each tube earlier placing any liquid within.
3. Draw tabular array 2 in your lab notebook and utilise it to tape your data.

Results

Table 2. Calculating Accuracy for Micropipetting

Tube #	Weight of tube (g)	Weight of tube + dye (g)	Theoretical weight of dye	Bodily weight of dye	% Error
1
two
3

4. Evangelize the volumes indicated in Table 3 into each of the iii labeled tubes.

Table 3. Volumes to be Pipetted into each Tube

Tube #	Micropipette	Reddish Dye (µl)	Blueish Dye (µl)	Green Dye (µl)
1	P1000	210	435	332
2	P200	110	153	67
3	P20	fifteen	17	10

5. Counterbalance each tube after pipetting.
6. Make up one's mind the theoretical weight of the dye using the data almost the weight of a mL of the dye solution at room temperature provided by your instructor.
7. Decide the % error for each tube.

Conclusion

Based on your data comment on the following in your lab notebook:

1. Which micropipette gave the most precise measurement?
2. Which micropipette gave the well-nigh accurate measurement?
3. What may accept contributed to higher percent errors?

Study Questions

1. Convert the following:
   • 345 mL = __________________ µl
   • 0.54 mL = _________________ µl
   • five.two 50 = ________________ mL
2. Which micropipette would yous choose to measure 550µl? 17µl? 167µl?
3. Make iii suggestions that other biotechnologists can utilise to amend micropipetting accuracy.
4. Assuming that the density of water is 1 gram per milliliter, how much should 550 µL of water counterbalance?
   • 17 µL of water?
   • 167 µL of water?
5. What is the formula to calculate percent fault?
6. What is the maximum book you tin set up for each micropipette (P-one thousand, P-200, P-twenty)?

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Source: https://bio.libretexts.org/Bookshelves/Biotechnology/Lab_Manual:_Introduction_to_Biotechnology/01:_Techniques/1.03:_Micropipetting

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