Team:UNITN-Trento/Notebook/Labposts/07/74

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{"date" : "2013-07-30","author" : "gabriele","title" : "pLAC to SAMsynthetase","content" : "<html>I digested pSB1C3+pLac with PstI-HF and SpeI and SAMsynthetase (PCR insert) with PstI-HF and XbaI, using the 'short' protocol (1.5 &micro;g of insert, 1 &micro;g of plasmid, for 2h at 37&deg;C). Then I treated as usual the plasmid with SAP and the insert with DpNI and purified them. The yields of the purification was too low, I was able to perform only Ctrl, 1:1 and 1:2 ligations.</html>","tags" : "SAMsynthetase-pLac"}
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"date":"2013-07-01",
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"author":"viola",
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"title":"<html>PCR of the double terminator BBa_B0015</html>",
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"content":"<html>Today i performed two PCR of the terminators both from the template in pSB1AK3 and from pSB1C£ following this quantities: <table> <tr> <th></th> <th>B0015 in pSB1AK3 <br/> 219,4 ng/ µl </th> <th>B0015 in pSB1C3 <br/> 413,3 ng/ µl </th></tr> <tr> <td>H20</td> <td>33.3 µl </td> <td>33.3 µl</td> </tr> <tr> <td>DNA</td> <td>0.5 µl</td> <td>0.5 µl</td> </tr> <tr> <td>DNTPs</td> <td>1 µl</td> <td>1 µl</td> </tr> <tr> <td>pref. FW</td> <td>2.5 µl</td> <td>2.5 µl</td> </tr> <tr> <td>suff. RV</td> <td>2.5 µl</td> <td>2.5 µl</td> </tr> <tr> <td>BUFFER</td> <td>10 µl</td> <td>10 µl</td> </tr> <tr> <td>Phusion pol.</td> <td>0.5 µl</td> <td>0.5 µl</td> </tr> </table> </html>}}</html>" I loaded the samples without dye and with 30% glycerol and this is the picture of the gel :</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/3/37/Tn-2013_Corsa20minPCR_B0015.jpg\" width=\"450px\" /></center></html>}}<html> on the left of the central 100bp ladder ther is B0015 in psb1ak3 and on the right there is B0015 in psb1c3.,
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"tags":"EFE"
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}
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Latest revision as of 10:49, 3 October 2013

{"date" : "2013-07-30","author" : "gabriele","title" : "pLAC to SAMsynthetase","content" : "I digested pSB1C3+pLac with PstI-HF and SpeI and SAMsynthetase (PCR insert) with PstI-HF and XbaI, using the 'short' protocol (1.5 µg of insert, 1 µg of plasmid, for 2h at 37°C). Then I treated as usual the plasmid with SAP and the insert with DpNI and purified them. The yields of the purification was too low, I was able to perform only Ctrl, 1:1 and 1:2 ligations.","tags" : "SAMsynthetase-pLac"}

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