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Cancer Borealis Saline

Salt (FW)Conc (mM)g/Lg/4Lg/6Lg/9Lg/11L
KCl (74.56)11.00.823.284.927.389.02
NaCl (58.44)440.025.67102.7154.28231282.85
CaCl2.2H2O (147.02)13.01.917.6411.4717.1921.02
MgCl2.6H2O (203.31)26.05.2921.1531.7247.6158.15
Trizma base (121.14)11.21.56.0913.514.92
Maleic acid (116.07)5.10.622.483.725.586.51
Hepes (instead of Tris+Maleate) (283.3)10.02.83 17.00 31.16

-pH should be between 7.4-7.5 (adjust with maleic acid and trizma base)

-throw out if not between 7.3-7.6 on first reading w/o adjustments

Cancer borealis High [K+] Saline Recipes

Chemicalg/6Lg/9Lg/L
NaCl154.00231.0025.67
MgCl2*H2O31.7447.615.29
CaCl2*H2O11.4617.191.91
KClsee chartssee chartssee charts
Trizma Base8.6012.901.43
Maleic Acid3.765.680.63

 

[K+] Varied Solutions

 
Amount of KCl to Add for: (in grams)
Volume of Saline0.5x1x1.5x2x2.5x3x
1L0.410.821.231.642.052.46
4L1.643.284.926.568.209.84
6L2.464.927.389.8412.3014.76
9L3.697.3811.0714.7618.4522.14

Panline Saline

Salt (FW)Conc (mM)g/Lg/4Lg/6Lg/9Lg/11L
KCl (74.56)11.00.823.284.928.559.02
NaCl (58.44)440.025.67102.7154.28252.0282.85
CaCl2 .2H2O (147.02)13.01.917.6411.4718.0921.02
MgSO4 anhydrous20.442.469.8414.7622.14 
Na2SO4 anhydrous3.910.552.23.34.45 
Hepes (hemisodium salt)5.02.499.9714.9622.44 

-pH should be at 7.45

-adjust with HCl and NaOH

Supplement Solution (to mix 50:50 with L-15 culture medium):

Salt (FW)Conc (mM)g/Lg/6Lg/11L
KCl (74.56)16.401.220.610.305
NaCl (58.44)743.6643.4621.7310.865
CaCl2.2H2O (147.02)24.743.641.820.909
MgCl2.6H20 (203.31)50.2010.215.102.552
Hepes (283.3)10.02.831.420.708

Zero Ca++ / zero Mg++ Dissociation Solution

Salt (FW)Conc (mM)g/Lg/6Lg/11L
KCl (74.56)11.00.820.410.205
NaCl (58.44)440.025.7112.866.43
Hepes (283.3)10.02.831.420.708

Osmolarity = 1030 mOsm

Saline is made regularly by the technician and the undergraduates, to keep at least three 6L flasks of saline ready at all times. Some days/weeks a lot of saline gets used and we may run out. In this case people can pitch in and make saline whenever they need.

Long-Term Cancer Preservation Saline

For 6L Erlenmeyers: Volume of MilliQ water: 5958ml 
After it is mixed, pH is tested and then it is vacuum filtered. 

 

Salt (FW)Conc (mM)g/Lg/4Lg/6Lg/9Lg/11L
KCl (74.56)11.00.823.284.927.389.02
NaCl (58.44)440.025.67102.7154.28231282.85
CaCl2.2H2O (147.02)13.01.917.6411.4717.1921.02
MgCl2.6H2O (203.31)26.05.2921.1531.7247.6158.15
Trizma base (121.14)11.21.56.09 (8.6)13.514.92
Maleic acid (116.07)5.10.622.483.725.586.51
Dextrose (180.16)5.551.004.006.009.0011.0
Fungizone   12ml  
Antibiotic (Pen Strep)   30ml  

 

Large Stock Cancer Saline Procedure

  1. Take used 9L saline container, remove the label, and rinse out three times with hot water and three times with deionized water: don’t use any soap or detergent.
  2. Bring the container to the bench with scales.
  3. Insert stirrer into the container.
  4. Obtain ‘Saline Mixing Checklist’ – start checking off items as you go.
  5. Add chemicals according to 9 L protocol on cards.
  6. Fill the container up to the line indicated: not the factory printed 6 Liter line, but the black permanent marker line with milliQ water.
  7. Set container on a magnetic stirrer. Allow salts to mix (at least a speed of 2-3) and dissolve for at least 30 minutes.
  8. Calibrate pH meter. Procedure for 2-point calibration of the pH meter can be found in the top drawer at the same bench right under the location of the meter.
  9. Take pH reading of saline, record value onto mixing checklist. 
  10. Adjust pH by adding acid or base to saline: which acid or base should be used depends on whether Cancer saline or Panline is being made: refer to note stuck on the first shelf at the same bench.
  11. Take pH reading, if it is within + 0.03 of 7.45 for saline and 7.4-7.5 for cancer, it is OK. Tape the checklist onto the flask: use red tape for Cancer saline, green tape for Panline. On the checklist record whether this is Cancer or Panline saline, final pH value, your name, and the date.
  12. Place flask in cold room for future use. Newer flasks are placed on the right side. 

More Saline Recipes!

Salines.pdf

Molarity Calculator

http://www.graphpad.com/quickcalcs/Molarityform.cfm

 

 

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