User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
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* 4°C, ∞ | * 4°C, ∞ | ||
{| {{table}} border="1" cellspacing="3" <!-- | {| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
Line 65: | Line 65: | ||
Dilutions: to final concentration of 20 fmol/μL (40 μL) | Dilutions: to final concentration of 20 fmol/μL (40 μL) | ||
{| {{table}} border="1" cellspacing="3" <!-- Dilution table --> | {| {{table}} border="1" cellspacing="3" <!-- Dilution table: x = length in bp ÷ measured ng/μL * 0.26 * 2 --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | DNA | | bgcolor=#cfcfcf | DNA | ||
| bgcolor=#cfcfcf | length (bp) | | bgcolor=#cfcfcf | length (bp) | ||
| bgcolor=#cfcfcf | ng/μL | | bgcolor=#cfcfcf | ng/μL | ||
| bgcolor=#cfcfcf | Vol. | | bgcolor=#cfcfcf | Vol. (x) | ||
| bgcolor=#cfcfcf | + dH<sub>2</sub>O | | bgcolor=#cfcfcf | + dH<sub>2</sub>O | ||
|- | |- | ||
| pSB1A3 || 2000 || 85.5 || | | pSB1A3 || 2000 || 85.5 || 12.2 || 27.8 | ||
|- | |- | ||
| H2B+his || 396 || 40.4 || | | H2B+his || 396 || 40.4 || 5.1 || 34.9 | ||
|- | |- | ||
| LOV || 429 || | | LOV || 429 || 18.6 || 12.0 || 28.0 | ||
|} | |} | ||
Golden Gate Reactions | Golden Gate Reactions | ||
# | # LOV + H2B+his + pSB1A3 | ||
# pSB1A3 | # pSB1A3 | ||
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| gg4 BL01 || 1.0 || --- | | gg4 BL01 || 1.0 || --- | ||
|- | |- | ||
| 10x ligase buffer || 1.0 || 1.0 | | 10x PRO ligase buffer || 1.0 || 1.0 | ||
|- | |- | ||
| NEB T4 lgase || 0.25 || 0.25 | | NEB T4 lgase || 0.25 || 0.25 |
Revision as of 18:21, 23 February 2013
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02/23/13
Type IIS Assembly, PCR: LOV-H2B+his
PCR
Thermal cycling
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