Team:Heidelberg/Templates/Del week15 L

==07-08-2013==
===Results Sequencing===


Send the following samples (amplified fragments of ''' ''D. acidovorans'' DSM-39 ''' and backbone pSB4K5 from the partsregistry) to GATC for sequencing:
* DelL (FS_14 to FS_15) with Primer FS_14
* DelL (FS_14 to FS_15) with Primer FS_15

<gallery>
File:L-FS 14.txt | Sequencing of DelL(FS_14 to FS_15) with Primer FS_14
File:L-FS 15.txt | Sequencing of DelL(FS_14 to FS_15) with Primer FS_15
</gallery>

===Amplification of DelL (FS_14 to FS_15; 1.4 kb)===

[[File:Heidelberg_20130807 22-13long(OP) 3x14-15(L)besch.png|150px|thumb|Amplifcation of DelOP and DelL (07.08); run at 100 V, 0.8 % gel (TAE)]]
[[File:Heidelberg_20130807 22-13long(OP) 3x14-15(L)cut.png|150px|thumb|Amplifcation of DelOP and DelL (07.08), cut]]


:'''Reaction'''
{| class="wikitable"
|-
! what !! µl   
|-
| ''D. acidovorans'' SPH-1 || 1
|-
| FS_14:  (1/10) || 2.5
|-
| FS_15:  (1/10) || 2.5
|-
| Phusion Master Mix || 25
|-
| DMSO || 2.5
|-
| dd H<sub>2</sub>O || 16.5
|}


:'''Conditions'''
{| class="wikitable"
|-
! colspan="3" | Biorad MyCycler
|-
! Cycles-PCR !!  temperature [°C] !! Time [s]
|-
| 1 || 98 || 10
|-
| rowspan="3"| 12 || 98 || 1
|-
| 66 ↓ 0.5 || 5
|-
| 72 || 42
|-
|rowspan="3"| 18 || 98 || 1
|-
| 63 || 5
|-
| 72 || 42
|-
| 1 ||72 || 12 min
|-
| 1 || 4 || inf
|}

'''Results:'''
* Amplification worked very well, we had bright bands on the right height.
* Bands were cut out and DNA purified using QIAquick Gel Extration Kit.

==08-08-2013==
===Concentration measurement of DelL===
{| class="wikitable"
|-
! Fragment !! Primer !! Date PCR !! Concentration
|-
| DelL  || FS14-FS15 || [[DelL#07-08-2013|07-08-2013]] || 51 ng/µl
|}

==09-08-2013==
===Amplification of DelL (FS_14 to FS_15; 1.4 kb)===

[[File:Heidelberg_20130809 2log 4xOPfromagarplate FS21-FS26 2xL 2log.png|150px|thumb|Amplifcation of DelOP (I), DelFG and DelL ; run at 100 V, 0.8 % gel (TAE)(09.08)]]
[[File:Heidelberg_20130809 2log 4xOPfromagarplate FS21-FS26 2xL cut.png|150px|thumb|Amplifcation of DelOP (I), DelFG and DelL ; run at 100 V, 0.8 % gel (TAE)(09.08)]]

:'''Reaction'''
{| class="wikitable"
|-
! what !! µl   
|-
| ''D. acidovorans'' || 1
|-
| FS_14:  (1/10) || 2.5
|-
| FS_15:  (1/10) || 2.5
|-
| Phusion Master Mix || 25
|-
| DMSO || 2.5
|-
| dd H<sub>2</sub>O || 16.5
|}

:'''Conditions'''
{| class="wikitable"
|-
! colspan="3" | Biorad MyCycler*
|-
! Cycles-PCR !!  temperature [°C] !! Time [s]
|-
| 1 || 98 || 10
|-
| rowspan="3"| 12 || 98 || 1
|-
| 66 ↓ 0.5 || 5
|-
| 72 || 42
|-
|rowspan="3"| 18 || 98 || 1
|-
| 63 || 5
|-
| 72 || 42
|-
| 1 ||72 || 12 min
|-
| 1 || 10 || inf
|}

'''Results:'''
* Amplification worked very well, we had bright bands on the right height.
* Bands were cut out and DNA purified using QIAquick Gel Extration Kit.