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| Hal Hawkington MD |
| Willing to Help Cloners |
| 2009.11.03 10:02:22 | |
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This is my favorite time of the year, when the MCC biotech students start their cloning projects.
Tags: Cloning Project Hits: 134 | Read more... |
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| Biotech-Billy |
| Good Luck Cloners |
| 2009.11.03 09:58:36 | |
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Oh my, is it already time for the cloning project? I see the menu is expanded this year. Should be good.
Best of luck, cloners.
Biotech Billy
Tags: Cloning Project | Biotech | MCC | BIO212AA Hits: 156 | Read more... |
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| Kikkert |
| proC and trp B gene sequences and primers |
| 2009.11.03 09:46:14 | |
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Scholars,
Visit the Documents section in the left hand menu and click on 212AA. In this folder are the sequences for and primers used to amplify the genes proC and trpB. Tags: Stan Kikkert | BIO212AA | Cloning Project Hits: 138 | Read more... |
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| Kikkert |
| And They're Off! |
| 2009.11.03 08:36:03 | |
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And They’re Off!
The students of BIO 212 AA have begun their cloning projects! Each student will PCR and subsequently clone the gene for T4 DNA Ligase as well as a biosynthetic gene from Agrobacterium radiobacter k84 into the plasmid vector pUC19. This semester we are looking at 2 A. radiobacter genes involved in proline (proC) and tryptophan (trpB) biosynthesis. Tags: Stan Kikkert | BIO212AA | Cloning Project Hits: 93 | Read more... |
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| Kikkert |
| E. coli lecture notes |
| 2009.08.31 08:04:41 | |
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Hi,
I just posted "E. coli Competent Cells" in the documents section. This lecture discusses E. coli cell structure, growth and the various strains available for making competent cells. (We will have this discussion on Monday while our cells are growing.) Tags: BIO212AA Hits: 126 | Read more... |
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| Kikkert |
| Latest Upgrade to Keep a Notebook |
| 2009.08.30 05:14:29 | |
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BIO 212AA Scholars,
I have just uploaded a more thorough description on How to Keep a Notebook. It is in the Document section under 21AA. Here's the link http://www.mccbiotechnology.net/bios/index.php?option=com_docman&task=cat_view&gid=107&Itemid=62 Tags: BIO 212 AA Hits: 128 | Read more... |
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| Kikkert |
| Welcome Fall 2009 |
| 2009.08.28 07:24:40 | |
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The doors to the lab have been thrown open and a new class of biotech recruits has walkwed hrough the door. Let's see if the website bbecomes a place where we can interact and talk molecular biology. Tags: BIO212AA Hits: 152 | Read more... |
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| kramer |
| T4 PCR didn't work |
| 2009.04.06 09:05:50 | |
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I think I'm the last one to get it done. Did it Friday. No joy! Saw the ladder but got NO BANDS in any other of the four lanes. My temps were 60C, 57C, 50.5C and 45C. My note book says I only put 6.25 uL of 10X PCR + Mg buffer in my coktail (1.25 uL per alliquotient across a 5X cocktail). Maybe that was my problem. Tags: Hits: 93 | Read more... |
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| kramer |
| Jenn's email question about subcloning |
| 2009.04.06 09:00:20 | |
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This web site is seriously weak...doesn't work with IE7...only Mozilla. Here is a cut-and-paste of an email conversation with Jenn about the subcloning exercise for BIO212AA S'09.
body {margin:8px} .tr-field {font:normal x-small arial} Jen, Sorry so late in replying. Daniel was walking through the same logic today with me, so I'll tell you the conclusions we came to... The pUC19 MCS is in the LacZ operon, so any changes there will make the gene inoperative. The cut is 23 nucleotides long so one would hope that if you stuck 23 "new nucleotides" in...well, at least there wouldn't be a frame shift mutation. I don't know that it's reasonable to expect proper expression of LacZ though...so it's anyone's guess whether blue-white selection will work any more. (I think not.) Re: polymerase synthesizing the genes...no, I don't think it understands content, but if you have nonsense nucleotide sequences, the synthesized protein won't work (reference blue-white selection discussion above). Soooo...yeh. I think you're right. If you successfully clone something else in there, LacZ won't work anymore. I'm going to cut-and-paste this discussion into the bio-blog-o-sphere (has the language really denegrated to THAT?) Peace, _ C On Fri, Apr 3, 2009 at 1:06 PM, jennifer kistner <
This e-mail address is being protected from spambots. You need JavaScript enabled to view it
> wrote:
Tags: BIO212AA subcloning pUC19 LacZ
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| Kikkert |
| Ghosts of the Past? |
| 2009.04.03 12:07:19 | |
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Are ghosts of the past rising?
First mass PCR attempt on T4 DNA Ligase gene nearly a complete failure. A great chance to trouble shoot, however. Hits: 126 | Read more... |
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 londonundergroundAZ
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| Good Luck |
| 2009.04.03 06:28:16 | |
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To the students of BIO212AA, as you set sail on your quest to successful clone a gene, I wish you all the luck in the world; you are going to need it. For those of you wit facebook, check the MCCBiotechClub Page, where someone has been posting some links you may or may not find useful. They are not the ultimate answer that you seek, but they may asisst you on your way. Mcc Biotech's Profile  Tags: BIO212AA | PCR Pirates of Penzance | Cloning Project Hits: 124 | Read more... |
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| kramer |
| BIO212AA-S2009: pMal-P2X digest looks like pUC-18 |
| 2009.03.05 06:07:34 | |
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re: digest of Mini-Prep-II pMal-P2X plasmid 27-Feb-2009 My pMal came out looking like pUC-18/19 with a monomer between 2kb and 3kb and clear dimer, trimer and quatramer bands appearing on the gel. Dan K got exactly the same results. We did not use the same DNA, did not share plates...anything. Two separate experiments. I'm wondering if what we thought was pMal was actually pUC? Tonights digest is only laid out for 5 lanes. I'm thinking about tossing another lane of pMal on tonights gel? Did anyone else get readable pMal on that last digest? Mark? Jenn? Does anyone else want to run some pMal tonight? -jak Tags:
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| londonundergroundAZ
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| D-Day |
| 2008.12.17 12:45:16 | |
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Seasickness and fear As the beaches draw near. Overhead artillery booms Illuminating the Normandy morning with the first flashes of battle. Soldiers in landing craft, donning their gear Saying final prayers, as bullets pierce the air announcing the enemy.
Landing craft doors slap the angry surf, Men jump into too deep waters, some carried by the weight of their loads or murderous fire, to the bosom of Neptune.
Others bravely wade to shore, Amid the ordinance, obstacles, blood and bodies of their comrades. Then rush to meagre shelter by the sea wall, As the officer shouts that only the dead and the soon to be dead will remain on these beaches under fire.
The armada of a thousand ships at dawn grow fear and stubborn determination in the the German defenders, And hope in the hearts of French civilians. Their resistance emerges into the light, as they pray these liberators shall prevail against their oppressors.
D-Day, December 17 2008. The beginning of the end of the greatest conflict in history, The freedom we now enjoy born with from this day’s terrible labour, Leaving debts to the soldiers living and dead That can never be repaid. Except that we are thankful, and promise never to forget their great sacrifices. Tags: D-Day | BIO212AA | biotechnology Hits: 114 | Read more... |
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| Rbarlow83 |
| Last lab tomorrow... |
| 2008.12.10 10:18:38 | |
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Tomorrow is my last chance to look for my T4-pUC18 clone. I have a feeling that it could be in there... My voodoo-dar is ringing off the hook. I also have some idea of what I want to keep doing if I don't find a clone. Be that as it may, those experiments will have to wait. Perhaps I'll fax an unnamed Englishman my protocols with the intent of having them carried out in secret... I feel as though I got into MCC biotechnology at the beginning of an beautiful era... like the times of Pax Romana or John Candy's movie career (both of equal importance and excellence in terms of splendor) But now... it is over. All over for RB. :( Hopefully I'll get that clone so I can tell my friends who all love to listen to me talk about science. :) There are two lies in that last sentence. See if you can figure them out! Tags:
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| Rbarlow83 |
| Calling all cars! Calling all cars! |
| 2008.12.06 12:37:41 | |
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Lets meet up this weekend for 212AA studies! Anyone! Everyone! But not teachers! We will push forward, using each others knowledge and resources as a means for intellectual growth. This ain't no ASU! Grab your notes and prepare to attack before like you've attacked! A great someone once said "don't let the wave hit you... you hit the wave..." If one human can do it, why can't we? Call me. Lets meet up. I just relized I have no phone minutes... email me... RB Sat: Any Sun: Any Hiya! Tags:
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| biotech |
| Final stretch |
| 2008.12.05 01:31:43 | |
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With time running out we are down to sticky end AP strategy. With the last transformations I did I really could not conclude whether I created any clones but my transformation protocol and spreading technique was pretty good. I have already done some ligation with sticky end with AP and will transform them tomorrow. Hopefully we will get some results early next week. Meanwhile it is interesting to see the lab with all of us at different stages of the cloning projects. There is competition & rivalry in the air. There is intimidation and suddenly from being classmates we have become individuals trying to be the one to get the 1st clone. These last couple of weeks from being totally clueless about bringing all the ideas and concepts we have learnt together into the cloning project the light is shinning at the end of the tunnel. Whether I get the clone or not I myself have transformed into a person with better understanding. Ultimately the fact is to clone “luck” has to favor you. So let us see who gets Lucky!!! All the Best Tags: Hits: 100 | Read more... |
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| Rbarlow83 |
| New Ligation... |
| 2008.12.03 12:06:05 | |
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I never got around to doing my ligation on Dec. 1. The vector I was using wasn't concentrated enough, so I redid my puc18 digest, but used more DNA. So... tomorrow I'm going to combine my T4 and puc18 in a ligation reaction... I'm really excited because I think it's going to work. I've put a lot of time (about 4 hours tonight) and thought into the protocol I'm running tomorrow... so my hopes are high. Soon we will do the ligation interpretive dance!
RB Tags: Hits: 85 | Read more... |
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| londonundergroundAZ
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| Crushmode |
| 2008.12.03 10:21:09 | |
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Time is running out. BIO212AA is like a circus. Tags: has anyone seen the PCR Pirate? | londonunderground | UK | crushmode | cloning | island | Biotech | BIO212AA | LeuS
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| Rbarlow83 |
| Vector Dephosphorylation Protocol |
| 2008.12.01 10:45:40 | |
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For Dec. 1 I'm going to dephosphorylate my pUC18 vector to get it ready for ligation with the T4 insert... From NEB:
Anyone want to join me? Anyone have any thoughts on this?
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| Biotech-Billy |
| Happy Thanksgiving from Biotech Billy! |
| 2008.11.29 07:14:35 | |
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Hey BioTechfriques, Happy Thanksgiving!
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