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arniekyo

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С°×3521

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scelab(½ð±Ò+2):ÈÈÐijæÓÑ£¬»¶Ó­¶àÀ´½»Á÷~~~:tiger06::tiger28::tiger23: 2010-04-16 22:00
arniekyo(½ð±Ò+1): 2010-08-15 14:56:38
ÒýÓûØÌû:
Originally posted by arniekyo at 2010-04-16 12:55:15:
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[ Last edited by С°×3521 on 2010-4-16 at 14:52 ]
2Â¥2010-04-16 14:50:31
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jingcp200320

Òø³æ (СÓÐÃûÆø)

¡ï ¡ï ¡ï
scelab(½ð±Ò+3):ÈÈÐijæÓÑ£¬»¶Ó­¶àÀ´½»Á÷~~~:tiger06::tiger28::tiger23: 2010-04-16 22:00
arniekyo(½ð±Ò+1): 2010-04-17 19:42
arniekyo(½ð±Ò+1): 2010-04-18 14:57
arniekyo(½ð±Ò+1): 2010-06-18 09:34:05
Recipes
Buffer Stock
Solutions (10X)
To prepare the buffer solutions described below, you need to prepare sodium
phosphate stock solutions:
Stock Solution A (10X)
200 mM sodium phosphate, monobasic (NaH2PO4)
5 M NaCl
Dissolve 27.6 g sodium phosphate, monobasic (NaH2PO4) and 292.9 g NaCl in
900 ml of deionized water. Mix well and adjust the volume to 1 L with
deionized water. Store solution at room temperature.
Stock Solution B (10X)
200 mM sodium phosphate, dibasic (Na2HPO4)
5 M NaCl
Dissolve 28.4 g sodium phosphate, dibasic (Na2HPO4) and 292.9 g of NaCl in
900 ml of deionized water. Mix well and adjust the volume to 1 L with
deionized water. Store solution at room temperature.
5X Native
Purification Buffer
250 mM NaH2PO4, pH 8.0
2.5 M NaCl
Prepare 200 ml solution as follows:
1. To 180 ml deionized water, add
Sodium phosphate, monobasic 7 g
NaCl 29.2 g
2. Mix well and adjust the pH with NaOH to pH 8.0.
3. Bring the final volume to 200 ml with water.
4. Store buffer at room temperature.
3 M Imidazole
pH 6.0
3 M Imidazole
500 mM NaCl
20 mM Sodium Phosphate Buffer, pH 6.0
Prepare 100 ml solution as follows:
2. To 90 ml deionized water, add
Imidazole 20.6 g
Stock Solution A (10X) 8.77 ml
Stock Solution B (10X) 1.23 ml
3. Mix well and adjust the pH to 6.0 with HCl or NaOH as necessary.
4. Bring the final volume to 100 ml with water. If the solution forms a
precipitate, heat solution until the precipitate dissolves.
5. Store buffer at room temperature.
Continued on next page
18
Recipes, Continued
Guanidinium Lysis
Buffer
6 M Guanidine Hydrochloride
20 mM Sodium Phosphate, pH 7.8
500 mM NaCl
Prepare 100 ml solution as follows:
1. To 90 ml deionized water, add
Stock Solution A (10X) 0.58 ml
Stock Solution B (10X) 9.42 ml
Guanidine Hydrochloride 57.3 g
2. Stir the solution until completely dissolved. Adjust the pH to 7.8 using
1 N NaOH or 1 N HCl.
3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 ¦Ìm
filter (autoclaving the solution will alter the pH of the buffer).
4. Store buffer at room temperature.
Denaturing Binding
Buffer
8 M Urea
20 mM Sodium Phosphate pH 7.8
500 mM NaCl
Prepare 100 ml solution as follows:
1. To 90 ml deionized water, add
Stock Solution A (10X) 0.58 ml
Stock Solution B (10X) 9.42 ml
Urea 48.1g
2. Stir the solution with gentle heating (50-60¡ãC, do not overheat) until
completely dissolved. When cooled to room temperature, adjust the pH
to 7.8 using 1 N NaOH or 1 N HCl.
3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 ¦Ìm
filter (autoclaving the solution will alter the pH of the buffer).
4. Store buffer at room temperature.
Continued on next page
19
Recipes, Continued
Denaturing Wash
Buffer
8 M Urea
20 mM Sodium Phosphate, pH 6.0
500 mM NaCl
Prepare 100 ml solution as follows:
1. To 90 ml deionized water, add
Stock Solution A (10X) 7.38 ml
Stock Solution B (10X) 2.62 ml
Urea 48.1g
2. Stir the solution with gentle heating (50-60¡ãC, do not overheat) until
completely dissolved. Adjust the pH to 6.0 using 1 N NaOH or 1 N HCl.
3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 ¦Ìm
filter (autoclaving the solution will alter the pH of the buffer).
4. Store buffer at room temperature.
Denaturing Elution
Buffer
8 M Urea
20 mM Sodium Phosphate, pH 4.0
500 mM NaCl
Prepare 100 ml as follows:
1. To 90 ml deionized water, add
Stock Solution A (10X) 10 ml
Urea 48.1g
2. Stir the solution with gentle heating (50-60¡ãC, do not overheat) until
completely dissolved. Adjust the pH to 4.0 using 1 N NaOH or 1 N HCl.
3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 ¦Ìm
filter (autoclaving the solution will alter the pH of the buffer).
4. Store buffer at room temperature.
3Â¥2010-04-16 15:57:03
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû

jingcp200320

Òø³æ (СÓÐÃûÆø)

arniekyo(½ð±Ò+2): 2010-08-15 14:56:49
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4Â¥2010-04-16 15:57:28
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