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| How much agarose is needed in making an 0.8% agarose gel with a final volume of mL? |
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Definition
| weight out 0.4g of agarose. add to a flask. add 50mL TAE or TBE |
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| How much agarose is needed in making an 1% agarose gel with a final volume of mL? |
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Definition
| weight out 0.5g of agarose. add to a flask. add 50mL TAE or TBE |
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| How much agarose is needed in making an 0.5% agarose gel with a final volume of mL? |
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Definition
| weight out 0.25g of agarose. add to a flask. add 50mL TAE or TBE |
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| How much agarose is needed in making an 1.5% agarose gel with a final volume of mL? |
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Definition
| weight out 0.75g of agarose. add to a flask. add 50mL TAE or TBE |
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Term
| You want to make a 2% agarose gel. The stock solution in the cabinet is 50X TAE and you want a final concentration of 1X TAE. You need 50 mL to form the gel. You also want to add ethidium bromide to the gel. The stock solution of EtBr is 10mg/ml but you only need 0.5 micograms/ml. Show you math and describe how you make the gel. |
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Definition
buffer = 1mL TAE, EtBr = 0.0025mL = 2.5 micro liters, Agarose = 1g agarose
procedure:
1. make 1X TAE by adding 1mL of stock 50mL TAE to a cylinder and then add 49mL of DiH2O
2. add 1g agarose to flask
3. pour in the 50mL 1X TAE and stir
4. heat in microwave at 30 second intervals
5. when solution is clear, it is done
6. when solution has cooled slightly, add 2.5 microliters EtBr
7. Mix solution well
8. pour into a gel cassette and add comb |
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Term
| you have a 50X TBE stock solution and you need 50mL of 1X TBE |
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Definition
| add 1ml of 50X TBE and 49mL of diH2O |
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| you have a 50X TBE stock solution and you need 50mL of 5X TBE |
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Definition
| add 5ml of 50X TBE and 45mL of diH2O |
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| you have a 50X TBE stock solution and you need 50mL of 15X TBE |
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Definition
| add 15ml of 50X TBE and 35mL of diH2O |
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| you have a 50X TBE stock solution and you need 50mL of 25X TBE |
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Definition
| add 25mL of 50X TBE and 25mL of diH2O |
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| how much solute is required to prepare a 100mL solution of NaCl 2.5M (MW = 58.44 g/mol) |
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Definition
g = MW x L x M
Weight out 14.16g NaCl. Add 100mL diH2O, Stir solution until blended |
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Term
| how would you make 300mL of a 0.8M solution of tris buffer (MW = 121.1 g/mol) |
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Definition
g = MW x L x M
weight out 29.06g of tris. add to a 500mL flask. add diH20 until you reach 300mL and blend solution |
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| You need 10% chelex in your lysis buffer, total volume is 1L how much of chelex would you have to add? |
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Definition
| % = g/mL x = 100g of chelex |
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| need an 0.8% agarose gel, stock solution is 50X TAE you want 1X TAE, stock EtBr is 10mg/ml but you want 0.5 microgram/ml. you need 80mL total to form the gel. |
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Definition
buffer = v1c1=v2c2 = 1.5 mL 50X TAE
agarose = g/mL = x/mL = 0.64g agarose
EtBr = c1v1 = c2v2 = 0.004 mL = 4 microliters EtBr |
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Term
| You need 25 mL of a 1:1000 dilution of primary antibody in blocking solution |
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Definition
multiply amount needed by dilution factor (1/1000) = 25 microliters
25 microliters of primary antibody was added to 25mL of blocking solution and incubate |
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Term
| You need 25 mL of a 2:1000 dilution of secondary antibody in blocking solution |
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Definition
25 mL x 2/1000 = 50/1000 = 0.05 mL = 50 microliters
50 microliters of secondary antibody was added to 25 mL of blocking solution and incubated |
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| What is the range of the P-1000 |
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| What is the range of the P-10 |
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| You have a 0.25 M solution of Na2EDTA on the shelf. You want to make 100ml of a 1mM solution for use in an experiment. How would you dilute the stock? |
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Definition
v1c1 = v2c2 = x 250 = 100 (1) = 0.40mL
withdraw 0.40mL of the 0.25M solution of Na2EDTA. Add to flask. add diH20 until solution reaches 100mL in the flask |
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| How many grams of NaCl would you need to make a 500mL of a 4M NaCl solution? |
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Definition
grams/molecular weight / volume = molarity
grams/58.44 / 0.5L = 4M
grams = 116.88g
place a dish on a balance. tare the scale. weight out 116.88g of NaCl onto the dish. add to flask. add dih20 until solution reaches 500mL in the flask |
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Term
| We have 38% stock acrylamide solution and 2% bis-acrylamide. How many grams of each reagent would you need to make up a 200mL solution? |
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Definition
38g x 2 = 76g of acrylamide. 2g x 2 = 4g bis-acrylamide. (x 2 because need 200mL)
place a dish on a scale. tare the scale. weight out 76g acrylamide. add to flask. add another dish and tare the scale. weight out 4g of bis-acrylamide. add to flash. add buffer until you reach 200mL in the flask to make a 200mL stock solution |
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Term
| you need to make a 10% acrylamide gel that requires 80mL total gel solution. your stock acrylamide is 40%. how much of the stock acrylamide would you use and how much buffer? |
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Definition
v1c1=c2c2, 20mL
withdraw 20mL of the 40% stock acrylamide. add to flask.
80-20 = 60mL, 60mL buffer to the 20mL of stock acrylamide until you reach 80mL (final volume). |
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| How much stock acrylamide would you need to make an 8% gel? from 40% stock |
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Definition
v1c1=v2c2 = 16mL
withdraw 16mL of the 40% stock acrylamide. add to flash.
80-16 = 64mL, add 64 mL to the 16mL of stock acrylamide until you reach 80mL |
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Term
| you want to make up 250uL mixture of 4 dNTPs, each a final concentration of 1mM. The stock concentration of the individual dNTPs is 10mM. How would you prepare the mixture in solution? |
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Definition
| v1c1=v2c2, 25uL x 4 = 100uL total, then add 150uL to get to 250uL |
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Term
| How would you prepare 500mL of a 10X TBE solution (Tris-Boric acid-Na2EDTA). 10X final concentrations: 1.0 tris (MW=121), 0.89M boric acid (mw=61.8), 0.01M Na2EDTA (mw= 372) |
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Definition
(grams/mw)/vol = M, g = M (mw)(v)
tris = M mw L = 60.5g
boric acid = 27.5g
Na2EDTA = 1.86g
add 60.5g of tris, 27.5g of boric acid, 1.86g of na2EDTA to a large beaker. Add diH20 until you reach 500mL. Stir solution to mix |
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Term
| prepare 200mL of a 0.25M KCl solution. you have 2M KCl stock. |
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Definition
| v1c1=v2c2= 25mL, add 25mL of 2M KCl stock solution to a flask. Add dih20 until the total volume reaches 200mL |
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Term
| 40mL of a 1.5% agarose gel in buffer? |
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Definition
% =g/mL = g=%mL=0.6
add 0.6g agarose to a flask, add buffer until you reach 40mL |
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Term
| How would you prepare 200mL of a stock 10% SDS solution in water? |
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Definition
%=g/ml, g=%ml=20g
add 20g SDS to a flask. Add dih20 until the flask is filled to 200mL |
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Term
| PCR master mix is 24uL and you add 1uL of the primer set, stock concentration is 10uM. What is the final concentration of the primers in the master mix? |
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Definition
v1c1=v2c2= 0.4uM
Final concentration of the primers in the master mix is 0.4uM |
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Term
| concentration of protein sample is 35.8mg/ml. want to load 100ug of protein on a 12% poly-acrylamide SDS gel, how much volume of protein sample would you mix with 2X loading solution to achieve 100ug? |
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Definition
35.8mg=358000ug and 1ml=1000uL
35800/1000 = 100/x x=2.8ul of protein sample is mixed with the loading solution |
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| 670 uL or 0.67 mL on P-1000 |
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Definition
| Moles of solute/L of solution |
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Definition
| 1 g mw of solute per liter of solution |
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Term
| if a 1% gel is 1g agarose per 100mL of buffer, how many grams of agarose will you need to make a small agarose? |
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Definition
| 1g/100ml = x/50ml, x= 50x1/100 = 0.50g of agarose |
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Term
| You have 10X stock solution of TAE buffer. you need 100mL of 1X TAE |
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Definition
| v1c1=v2c2, 1mL of 10X TAE to 99mL of diH20 |
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Term
| you want to make a 1% agarose gel and we need 50mL total to make a small gel |
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Definition
1g/100ml = x/50ml = 0.5g
add 0.5g of agarose to a 250mL flash add 1X TAE until you reach 50mL |
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Term
| EtBr stock is 10mg/ml and you need 5ug/ml |
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Definition
| v1c1=v2c2, 2.5uL of EtBr to gel solution |
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Term
| final concentration of Taq polymerase is to be 0.01 units/uL in a 50uL PCR. If the enzyme supplied if 5 units/uL, how much enzyme would you add to the reaction? |
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| SSC is a buffer in blot NP-40 is a detergent, you need 300mL of 0.2% NP-40 in 2X SSC. Stock for NP-40 is 100%, how much NP-40 do you need to make this solution? |
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