3.1.5: Double Digestion Supplies
- Page ID
- 67612
\( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\)
Double Digestion:
Enzymes and buffers are indicated from New England Biolabs.
|
Materials |
Amount Needed (Per Reaction) |
|
Distilled Water |
25 uL-(Sum of DNA+NEB+Enzyme1+Enzyme2) |
|
DNA Sample |
Determined by spectrophotometer (300ng/DNA concentration) |
|
10x NEBuffer 2.1 (Cat#: B7202S) |
2.5 uL |
|
Restriction Enzyme 1 (EcoRI [Cat#: R0101S]) or SpeI [Cat#: R0133S]) |
1 uL |
|
Restriction Enzyme 2 (XbaI [Cat#: R0145S] or PstI [Cat#: R0140S]) |
1 uL |
|
Water/Bead Bath (37C) |
1 (used by class) |
|
Heat Block (80C) |
1 (used by class) |
|
Microcentrifuge Tube |
1 |
|
Micropipette and Tips |
1-10uL, 2-20uL |
|
Microcentrifuge |
1 (used by class) |