Calendar/2011-3-28: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Alan6017518
>Alan6017518
Line 2: Line 2:
===Qubit Quantification===
===Qubit Quantification===
*dilute 25ul 200X dye with 4975ul buffer
*dilute 25ul 200X dye with 4975ul buffer
{| {{table}}
| Sample||Concentration in the Qubit||||µL used||Dilution||Sample Concentration||
|-
| ||||||||||||
|-
| 1||37.2||ng/mL||1||200||7.44||µg/mL
|-
| ||||||||||||
|-
| 2||103||ng/mL||1||200||20.6||µg/mL
|-
| ||||||||||||
|-
| 3||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 4||27||ng/mL||1||200||5.4||µg/mL
|-
| ||||||||||||
|-
| 5||47.8||ng/mL||1||200||9.56||µg/mL
|-
| ||||||||||||
|-
| 6||12.7||ng/mL||1||200||2.55||µg/mL
|-
| ||||||||||||
|-
| 7||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 8||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 9||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 10||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 11||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 12||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 13||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 14||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 15||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 16||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 17||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 18||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 19||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 20||Out Of Range||||||||||
|-
| ||||||||||||
|-
| 21||Out Of Range||||||||||
|}


===Tagmentation Reaction===
===Tagmentation Reaction===

Revision as of 21:08, 28 March 2011

Library Construction on sample 1-21 from Dr. Heller

Qubit Quantification

  • dilute 25ul 200X dye with 4975ul buffer
Sample Concentration in the Qubit µL used Dilution Sample Concentration
1 37.2 ng/mL 1 200 7.44 µg/mL
2 103 ng/mL 1 200 20.6 µg/mL
3 Out Of Range
4 27 ng/mL 1 200 5.4 µg/mL
5 47.8 ng/mL 1 200 9.56 µg/mL
6 12.7 ng/mL 1 200 2.55 µg/mL
7 Out Of Range
8 Out Of Range
9 Out Of Range
10 Out Of Range
11 Out Of Range
12 Out Of Range
13 Out Of Range
14 Out Of Range
15 Out Of Range
16 Out Of Range
17 Out Of Range
18 Out Of Range
19 Out Of Range
20 Out Of Range
21 Out Of Range

Tagmentation Reaction

  • Prior to assembling the reaction, briefly centrifuge the 5X Nextera Reaction Buffer and Nextera Enzyme Mix tubes to assure that the reagents are at the bottom of the tubes.
  • Assemble the following reaction components on ice, in the order listed:
x µl Nuclease-Free Water 
50 ng Target DNA (in T10E1 Buffer [10 mM Tris-HCl (pH 7.5), 1 mM EDTA])
4 µl 5X Nextera Reaction Buffer LMW or HMW (see Important Considerations, no. 1, p. 3)
1 µl Nextera Enzyme Mix (Illumina-compatible)
20 µl Total reaction volume