User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/11/20: Difference between revisions
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'''Digest (Fermentas FD)'''<br> | '''Digest (Fermentas FD)'''<br> | ||
# pT7CFE1-CHis, E/P | # pT7CFE1-CHis, E/P | ||
# KAH109, E/P | |||
# KAH111, E/P | |||
# KAH112, E/P | |||
# KAH115, E/P | |||
# KAH148, E/P | |||
# KAH150, E/P | |||
# KAH151, E/P | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | ||
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| <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u> | | <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u> | ||
|- | |- | ||
| 1. pT7CFE1-CHis (E/P) || --- || --- || --- | | 1. pT7CFE1-CHis (E/P) || 57.344 || 1.829 || --- | ||
|- | |||
| 2. KAH109, (E/P) || --- || 36.072 || 2.354 | |||
|- | |||
| 3. KAH111, (E/P) || --- || 15.003 || 2.238 | |||
|- | |||
| 4. KAH112, (E/P) || --- || 22.857 || 1.862 | |||
|- | |||
| 5. KAH115, (E/P) || --- || 18.312 || 2.096 | |||
|- | |||
| 6. KAH148, (E/P) || --- || 23.735 || 2.389 | |||
|- | |||
| 7. KAH150, (E/P) || --- || 19.822 || 1.99 | |||
|- | |||
| 8. KAH151, (E/P) || --- || 19.701 || 1.90 | |||
|} | |} | ||
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* Note: Use 50 ng of vector this time (instead of 20) | * Note: Use 50 ng of vector this time (instead of 20) | ||
* Note: Use a 3:1 ratio of insert to vector... x μL insert = 3 * (bp of insert/ bp of vector) * 50 ng vector / (concentration of insert ng/μL) | * Note: Use a 3:1 ratio of insert to vector... x μL insert = 3 * (bp of insert/ bp of vector) * 50 ng vector / (concentration of insert ng/μL) | ||
# BD-116 (KAH109) | |||
# BD-117 (KAH111) | |||
# BD-118 (KAH112) | |||
# BD-119 (KAH115) | |||
# BD-120 (KAH148) | |||
# BD-121 (KAH150) | |||
# BD-122 (KAH-151) | |||
# CTRL | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table --> | ||
| || 1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 | | || 1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 | ||
|- | |- | ||
| Insert DNA || | |Inser Name || KAH109 || KAH111 || KAH112 || KAH115 || KAH148 || KAH150 || KAH151 || N/A | ||
|- | |||
| Insert DNA || 1.3 || 3.12 || 2.05 || 2.12 || 1.7 || 2.03 || 2.05 || 0 | |||
|- | |- | ||
| Vector DNA || | | Vector DNA || 0.87 || 0.87 || 0.87 || 0.87 || 0.87 || 0.87 || 0.87 || 0.87 | ||
|- | |- | ||
| 2x lgn buf (Roche) || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 | | 2x lgn buf (Roche) || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 | ||
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| T4 ligase (NEB) || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 | | T4 ligase (NEB) || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 1.83 || 0.01 || 1.08 || 1.01 || 1.43 || 1.1 || 1.08 || 3.13 | ||
|- | |- | ||
| || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL | | || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL |
Revision as of 12:00, 26 November 2012
PcTF Subcloning in E. coli | <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> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
11/20/12Assemblies: Shown as Part/cuts/purified fragment size
Measure concentration(s)
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