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My Experimental World

Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. Saturday, July 31, 2010.

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My Experimental World | syahiedasha.blogspot.com Reviews

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Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. Saturday, July 31, 2010.

<META> KEYWORDS

1 my experimental world

2 progress 2

3 i already

4 reculture

5 and also the

6 purification was done

7 thus

8 i'm trying to

9 produce more protein

10 since

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My Experimental World | syahiedasha.blogspot.com Reviews

https://syahiedasha.blogspot.com

Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. Saturday, July 31, 2010.

INTERNAL PAGES

syahiedasha.blogspot.com

1

My Experimental World: ~What I have Done??~

http://www.syahiedasha.blogspot.com/2010/07/what-i-have-done.html

Saturday, July 24, 2010. What I have Done? I've got a clone called M15-pQEC1 (Escherichia coli M15 as a host, pQE-30 as expression vector that carried phaC1 gene). Then I continued with optimization of protein expression condition as below :. Culture was grown first at. 37°C, 180 rpm for 2 and a half hour until. Reach 0.5 -0.6. Midexponential phase), before induction of 0.01 mM IPTG. Extraction of expressed total protein and also solubility test was done in order to determine at. I found that induction at.

2

My Experimental World: ~Progress~

http://www.syahiedasha.blogspot.com/2010/07/progress.html

Saturday, July 31, 2010. Previously, I informed that induction at 28°C give more soluble form compared with other induction temperature. While M15-pQEC1/pGro7 give better result than M15-pQEC1. Thus, my attempt is to grow the cell at 28°C. Until it reach OD600 reach 0.5 -0.6. Midexponential phase),and also the induction was also at the same temperature. 2)Since there's a lot of contaminants, I've decided to do gel filtration (size -exclusion chromatography)to get purity of protein. Rerun the SDS-PAGE to ...

3

My Experimental World: July 2010

http://www.syahiedasha.blogspot.com/2010_07_01_archive.html

Saturday, July 31, 2010. Previously, I informed that induction at 28°C give more soluble form compared with other induction temperature. While M15-pQEC1/pGro7 give better result than M15-pQEC1. Thus, my attempt is to grow the cell at 28°C. Until it reach OD600 reach 0.5 -0.6. Midexponential phase),and also the induction was also at the same temperature. 2)Since there's a lot of contaminants, I've decided to do gel filtration (size -exclusion chromatography)to get purity of protein. Rerun the SDS-PAGE to ...

4

My Experimental World: August 2010

http://www.syahiedasha.blogspot.com/2010_08_01_archive.html

Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. Subscribe to: Posts (Atom).

5

My Experimental World: ~Progress 2~

http://www.syahiedasha.blogspot.com/2010/08/progress-2.html

Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. View my complete profile.

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adrian-protein.blogspot.com

Dimension: Insoluble protein?!

http://adrian-protein.blogspot.com/2010/08/insoluble-protein.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Wednesday, August 18, 2010. According to SDS P.A.G.E for solubility which I did on 18 AUG, the result doesn't looks good. Maybe it is better to subclone the stored plasmid to Rosetta-gami2 (DE3) and BL21 (DE3) , BL21 (DE3)/pLySs? At the same time, design primer and clone into pCold and pMAL.

adrian-protein.blogspot.com

Dimension: The definition of biodegradability

http://adrian-protein.blogspot.com/2010/12/definition-of-biodegradability.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Wednesday, December 22, 2010. The definition of biodegradability. Towards Common Ground –. Meeting Summary of the International Workshop on Biodegradability,. Annapolis, MD, USA, 1992. Such as composting, sewage treatment, denitrification, or anaerobic sludge treatment. Stuck @ dec 2010.

adrian-protein.blogspot.com

Dimension: August

http://adrian-protein.blogspot.com/2010/08/august.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Monday, August 16, 2010. Last 2 weeks screening for the culture of RG2 pET32 phaCcs between 2 culture from glycerol stock. As a result, one of the bacteria was contamination, didn't express protein phaC and didnt contain the correct plasmids. 17 AUG 2010 (Done). 1 Purification by TALON.

adrian-protein.blogspot.com

Dimension: Crystallization tutorial

http://adrian-protein.blogspot.com/2014/06/crystallization-tutorial.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Wednesday, June 4, 2014. This is a very simple tutorial about protein crystallization:. Http:/ xray.bmc.uu.se/ terese/tutorials.html. Bergfors, T., Editor. Protein Crystallization: Second Edition. 2009. International. University Line, La Jolla, California, 500 pp. KAS project by hani hadi.

adrian-protein.blogspot.com

Dimension: December 2010

http://adrian-protein.blogspot.com/2010_12_01_archive.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Wednesday, December 22, 2010. The definition of biodegradability. Towards Common Ground –. Meeting Summary of the International Workshop on Biodegradability,. Annapolis, MD, USA, 1992. Such as composting, sewage treatment, denitrification, or anaerobic sludge treatment. Stuck @ dec 2010.

adrian-protein.blogspot.com

Dimension: August 2010

http://adrian-protein.blogspot.com/2010_08_01_archive.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Sunday, August 22, 2010. This week will be a busy week starting from tomorrow. Maybe should say it is a busy month too. I do not want to waste my time anymore. So let's get to work. An amount of bacterial hosts and expression vectors appearing in my minds now. The NEB express - ER2523. As a ...

adrian-protein.blogspot.com

Dimension: Stuck @ dec 2010

http://adrian-protein.blogspot.com/2010/12/stuck-dec-2010.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Wednesday, December 8, 2010. Stuck @ dec 2010. E coli Rosettagami2(DE3) with pET32 harbouring pha. Problem 1: Expression level of PhaC is very low compare to previous batches. Less than 10% , will be very difficult to be purified). Problem 2: Almost 100% of the expressed PhaC is insoluble.

adrian-protein.blogspot.com

Dimension: July 2010

http://adrian-protein.blogspot.com/2010_07_01_archive.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Friday, July 30, 2010. Was trying to produce more protein from the 1.6L induced bacterial cultures. Purified with Affinity chromatography successfully. (Although lost an amount of protein due to my mistakes). Concentrated it with a protein concentrator since it has been diluted. The culture ...

adrian-protein.blogspot.com

Dimension: JUNE-JULY

http://adrian-protein.blogspot.com/2010/07/june-july.html

The X-ray study of proteins is sometimes regarded as an abstruse subject comprehensible only to specialists, but the basic ideas underlying our work are so simple that some physicists find them boring. (Perutz 1992). Friday, July 30, 2010. Was trying to produce more protein from the 1.6L induced bacterial cultures. Purified with Affinity chromatography successfully. (Although lost an amount of protein due to my mistakes). Concentrated it with a protein concentrator since it has been diluted.

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Monday, August 16, 2010. M15-pQEC1/pGro7 and M15-pQEC1 in 400 ml LB at. Induction was also at the same temperature. All the elution give approximately a band at around 60 kDa. Based on the standard curve.I managed to get about. 035 mg/ml of protein for. 02 mg/ml of protein for M15-pQEC1. I'm thinking of.if from 400 ml of culture, I get the amount shown above. Theoritically, I'll get higher concentration of protein from large amount of culture. From the 2.0 L bacterial cultures. Saturday, July 31, 2010.

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