Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/21: Difference between revisions
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'''Assemblies''' | '''Assemblies''' | ||
# H2B - X/S - 410 bp + pSB1A3 - X/S - 2000 bp | # NEB T4 ligase; H2B - X/S - 410 bp + pSB1A3 - X/S - 2000 bp | ||
# LOV - X/S - 461 bp + pSB1A3 - X/S - 2000 bp | # NEB T4 ligase; LOV - X/S - 461 bp + pSB1A3 - X/S - 2000 bp | ||
# (negtaive control) pSB1A3 - X/S - 2000 bp | # NEB T4 ligase; (negtaive control) pSB1A3 - X/S - 2000 bp | ||
# Roche T4 ligase; H2B - X/S - 410 bp + pSB1A3 - X/S - 2000 bp | |||
# Roche T4 ligase; LOV - X/S - 461 bp + pSB1A3 - X/S - 2000 bp | |||
# Roche T4 ligase; (negtaive control) pSB1A3 - X/S - 2000 bp | |||
* Digests: | * Digests: | ||
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'''Dephosphorylation''' | '''Dephosphorylation''' (11/23/12) | ||
> Dephosphorylation (Roche) | > Dephosphorylation (Roche) | ||
# pSB1A3 - X/S = | # pSB1A3 - X/S = 2000 bp | ||
{| class="wikitable" border="0" cellspacing="3" <!-- Dephos table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Dephos table --> | ||
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* Use 2x moles of H2B insert, relative to vector: xμL H2B insert = 3 * 410 bp H2B / 2000 bp pSB1A3 * 20 ng pSB1A3 / ### ng/μL H2B = '''### μL H2B''' | * Use 2x moles of H2B insert, relative to vector: xμL H2B insert = 3 * 410 bp H2B / 2000 bp pSB1A3 * 20 ng pSB1A3 / ### ng/μL H2B = '''### μL H2B''' | ||
* Use 2x moles of LOV insert, relative to vector: xμL LOV insert = 3 * 461 bp LOV / 2000 bp pSB1A3 * 20 ng pSB1A3 / ### ng/μL LOV = '''### μL LOV''' | * Use 2x moles of LOV insert, relative to vector: xμL LOV insert = 3 * 461 bp LOV / 2000 bp pSB1A3 * 20 ng pSB1A3 / ### ng/μL LOV = '''### μL LOV''' | ||
{| class="wikitable" width=400px | {| class="wikitable" width=400px |
Revision as of 14:32, 23 November 2012
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
11/21/12Results from yesterday's ligation
PCR program:
Assemblies
> Dephosphorylation (Roche)
Proceed directly to Transformation. See http://openwetware.org/wiki/Haynes:Assembly101
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