Team:TU Delft/29 July 2010 content

From 2010.igem.org

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The digestion products were [[Team:TU_Delft/protocols/ligation|ligated]] overnight:
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Revision as of 07:11, 2 August 2010

Lab work

Alkane Degradation

PCR Amplification

K398007A, K398008A, K398009A, K398010A, K398017A, K398018A, K398019A, B0015 were amplified with the universal primers G00100 and G00101. The product was put on 1% agarose gel:

Lane Description:

# Description Expected Length (bp) Primers Status Remarks
M1 SmartLadder n/a n/a n/a
1 PCR product of 007 1616 G00101 + G00101 ✔
2 PCR product of 008 473 G00101 + G00101 ✔
3 PCR product of 009 482 G00101 + G00101 ✔
4 PCR product of 010 1505 G00101 + G00101 ✔
5 PCR product of 017 1625 G00101 + G00101 ✔
6 PCR product of 018 1052 G00101 + G00101 ✔
7 PCR product of 019 1796 G00101 + G00101 ✔
8 PCR product of B0015 445 G00101 + G00101 ✔

The PCR products were subsequently digested:

# Sample Enzyme 1 Enzyme 2 Buffer BSA Needed fragment
1 40 μL PCR product 007 EcoRI SpeI 2 (BioLabs) ✓ ‘E–007–S’
2 40 μL PCR product 008 XbaI PstI 2 (BioLabs) ✓ ‘X–008-P’
3 40 μL PCR product 009 EcoRI SpeI 2 (BioLabs) ✓ ‘E–009–S’
4 40 μL PCR product 010 XbaI PstI 2 (BioLabs) ✓ ‘X–010-P’
5 40 μL PCR product 017 EcoRI SpeI 2 (BioLabs) ✓ ‘E–009–S’
6 40 μL PCR product 018 XbaI PstI 2 (BioLabs) ✓ ‘X–018-P’
7 40 μL PCR product 019 EcoRI SpeI 2 (BioLabs) ✓ ‘E–019-S’
8 40 μL PCR product B0015 XbaI PstI 2 (BioLabs) ✓ ‘X–B0015-P’

The digestion products were ligated overnight:

# BioBrick Fragment 1 Fragment 2 Final volume
1 4 μL ‘E–007–S’ 4 μL ‘X–00–P’
2 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
3 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
4 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
5 ? μL ‘E-Part1-S’ ? μL ‘X-Part2-P'
6 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
7 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
8 ? μL ‘E – Part – S’ ? μL ‘X – Part – P’
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