Berk2006-ConjugationTeam3: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
JCAnderson (talk | contribs) |
|||
(13 intermediate revisions by 2 users not shown) | |||
Line 1: | Line 1: | ||
==JL_11/29/06== | |||
# ran gel:<br> | |||
(Lane 1=1, lane 2=2... etc. Lane 13= ladder)<br> | |||
[[Image:JL112906-gel.jpg]] | |||
==JL_11/27/06== | |||
'''Tecan measurements''' <br> | |||
{| border="2" cellpadding="10" | |||
!width="15"|Culture | |||
!width="5"|Tecan measurement | |||
|- | |||
|''' TG1'''||42 | |||
|- | |||
|'''J01122'''||4 | |||
|- | |||
|'''J01022'''||2987 | |||
|- | |||
|'''J23090'''||44 | |||
|- | |||
|'''J23090-J23086'''||145 | |||
|- | |||
|'''J23093'''||55 | |||
|- | |||
|'''J23093-J23086'''||200 | |||
|} | |||
==JL_11/20/06== | |||
#made -80 of [lock 3n]J23093 | |||
#plated [lock 3n]J23093 | |||
#grew up [lock 3n]J23093 | |||
#organized boxes in freezer | |||
==JL_10/13/06== | |||
Conjugated EC100d with | |||
#J01064 db1K0 #1 | |||
#J01064 db1K0 #2 | |||
#J01064 db1K0 #3 | |||
#J01064 db1K0 #4 | |||
#J23015-J01064 | |||
on Tri Amp | |||
Also I made TriK and TriA plates | |||
==JL_10/09/06== | |||
#Mate J23016 with Ec100D (see if the trbC knockout transfers its | |||
plasmid, so plate on TriK) | |||
#Do genomic minipreps of J23087 (4 clones) | |||
#Mini and map J23086 (4 clones) | |||
#Mini and map J23088 (2 clones) | |||
==JL_09/29/06== | |||
#Preperative digest of pJ23006-J23006 (AlwnI/Spei, 2068 Long) | |||
#Preperative digest of psB1A2-J01003 (AlwnI/Xbai, 925 short) | |||
#4 Analytical digests(AvrII/SpeI) each of J23074 (want band at 23097) and J23075 (want band at 824) | |||
#grow up J23016 | |||
==JL_09/22/06== | |||
Today's to do list: | |||
#stock and mini pSB1AG0-J23078, map EcoRI/SpeI | |||
# grow up J23055 #1 for electro comp | |||
#grow up J23016 (trbC::Cm) for the knockout (later gen. mini and PCR with JL16/17) | |||
#Rlam/J23057 x pSB1A2-J01003 redo | |||
==JL_09/20/06== | |||
Today I... | |||
- mini-preped J23074 and J23075, and pSB1AKG0-b0015 | |||
- gen. mini and PCR J23055 | |||
- grew up Rlam/J23057 | |||
- analytical digest of J23074 and J23075 | |||
==[[User:JCAnderson|JCAnderson]] 22:35, 20 September 2006 (EDT)== | ==[[User:JCAnderson|JCAnderson]] 22:35, 20 September 2006 (EDT)== | ||
[[Image:JCA092006-gel.jpg|100px|right]] | |||
Four clones were spotted the other day, all 4 were ampS and camS. The JL3/4 PCRs were done on genomic preps today. Jen setup and sam ran PCRs of J23055 clones 1 and 2. Clone 1 looks correct, it was stocked and replated to make comp cells.<br> | Four clones were spotted the other day, all 4 were ampS and camS. The JL3/4 PCRs were done on genomic preps today. Jen setup and sam ran PCRs of J23055 clones 1 and 2. Clone 1 looks correct, it was stocked and replated to make comp cells.<br> | ||
==JL_09/18/06== | ==JL_09/18/06== |
Latest revision as of 14:26, 29 November 2006
JL_11/29/06
- ran gel:
(Lane 1=1, lane 2=2... etc. Lane 13= ladder)
JL_11/27/06
Tecan measurements
Culture | Tecan measurement |
---|---|
TG1 | 42 |
J01122 | 4 |
J01022 | 2987 |
J23090 | 44 |
J23090-J23086 | 145 |
J23093 | 55 |
J23093-J23086 | 200 |
JL_11/20/06
- made -80 of [lock 3n]J23093
- plated [lock 3n]J23093
- grew up [lock 3n]J23093
- organized boxes in freezer
JL_10/13/06
Conjugated EC100d with
- J01064 db1K0 #1
- J01064 db1K0 #2
- J01064 db1K0 #3
- J01064 db1K0 #4
- J23015-J01064
on Tri Amp
Also I made TriK and TriA plates
JL_10/09/06
- Mate J23016 with Ec100D (see if the trbC knockout transfers its
plasmid, so plate on TriK)
- Do genomic minipreps of J23087 (4 clones)
- Mini and map J23086 (4 clones)
- Mini and map J23088 (2 clones)
JL_09/29/06
- Preperative digest of pJ23006-J23006 (AlwnI/Spei, 2068 Long)
- Preperative digest of psB1A2-J01003 (AlwnI/Xbai, 925 short)
- 4 Analytical digests(AvrII/SpeI) each of J23074 (want band at 23097) and J23075 (want band at 824)
- grow up J23016
JL_09/22/06
Today's to do list:
- stock and mini pSB1AG0-J23078, map EcoRI/SpeI
- grow up J23055 #1 for electro comp
- grow up J23016 (trbC::Cm) for the knockout (later gen. mini and PCR with JL16/17)
- Rlam/J23057 x pSB1A2-J01003 redo
JL_09/20/06
Today I... - mini-preped J23074 and J23075, and pSB1AKG0-b0015 - gen. mini and PCR J23055 - grew up Rlam/J23057 - analytical digest of J23074 and J23075
JCAnderson 22:35, 20 September 2006 (EDT)
Four clones were spotted the other day, all 4 were ampS and camS. The JL3/4 PCRs were done on genomic preps today. Jen setup and sam ran PCRs of J23055 clones 1 and 2. Clone 1 looks correct, it was stocked and replated to make comp cells.
JL_09/18/06
Today I have to:
- Grow up J23074 and J23075 (4 clones each)
- Grow and spot J23055 (43 degrees J23015 x pCP20)
- Run PCR of J23069 with your traG 20mers