Maloof Lab:miniprep reagents: Difference between revisions
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(New page: '''Buffer 1 (Cell resuspension buffer''' 50mM Tris 10mM EDTA 100 ug/mL RNase A For 100 mL 5 mL 1M Tris pH8 2 mL 0.5M EDTA 1 mL 10mg/mL RNase A 92 mL water '''Buffer 2 (Cell lysis soluti...) |
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'''Buffer 1 (Cell resuspension buffer''' | '''Buffer 1 (Cell resuspension buffer)''' | ||
50mM Tris | 50mM Tris | ||
| Line 11: | Line 11: | ||
92 mL water | 92 mL water | ||
'''Buffer 2 (Cell lysis solution''' | '''Buffer 2 (Cell lysis solution)''' | ||
0.2M NaOH | 0.2M NaOH | ||
| Line 20: | Line 20: | ||
5 mL 20% SDS | 5 mL 20% SDS | ||
75 mL water | 75 mL water | ||
'''Buffer 3 (Neutralization Solution)''' | |||
4M guanidine hydrochloride | |||
0.5M sodium acetate | |||
For 100 mL | |||
2.3g guanidine hydrochloride | |||
4.9g potassium acetate | |||
pH to 4.2 with glacial acetic acid | |||
bring volume to 100 mL with water | |||
'''Wash buffer''' | |||
60mM potassium acetate | |||
8.3mM Tris-HCl (pH 7.5) | |||
40um EDTA (pH 8) | |||
60% ethanol | |||
For 100mL | |||
1.2 mL 5M potassium acetate | |||
0.83 mL 1M Tris (pH 7.5) | |||
8 ul 0.5M EDTA | |||
60 mL 100% ethanol | |||
38mL water | |||
ADD 170 mL 95% ethanol to the wash buffer before first use!! | |||
Revision as of 14:42, 21 October 2014
Buffer 1 (Cell resuspension buffer)
50mM Tris 10mM EDTA 100 ug/mL RNase A
For 100 mL 5 mL 1M Tris pH8 2 mL 0.5M EDTA 1 mL 10mg/mL RNase A 92 mL water
Buffer 2 (Cell lysis solution)
0.2M NaOH 1% SDA
For 100 mL 20 mL 1M NaOH 5 mL 20% SDS 75 mL water
Buffer 3 (Neutralization Solution)
4M guanidine hydrochloride 0.5M sodium acetate
For 100 mL 2.3g guanidine hydrochloride 4.9g potassium acetate pH to 4.2 with glacial acetic acid bring volume to 100 mL with water
Wash buffer
60mM potassium acetate 8.3mM Tris-HCl (pH 7.5) 40um EDTA (pH 8) 60% ethanol
For 100mL 1.2 mL 5M potassium acetate 0.83 mL 1M Tris (pH 7.5) 8 ul 0.5M EDTA 60 mL 100% ethanol 38mL water
ADD 170 mL 95% ethanol to the wash buffer before first use!!
