SlideShare a Scribd company logo

DNA & RNA extraction

•
•
Mohammed Bahgat Mohammed Sofyan
Mohammed Bahgat Mohammed Sofyan

DMBMS

Education
1 of 23
DNA,RNA extraction BY dr: Mohammed Bahgat Mohammed Sofyan Assistant lecturer of medical biochemistry, Faculty of medicine, Al-Azhar university (Assiut branch) 1
2
‫على‬ ‫التوكل‬ ‫هللا‬ 3
‫صورة‬ ‫كل‬ ‫على‬ ‫اضغط‬ ‫الشاش‬ ‫عرض‬ ‫وضع‬ ‫في‬ ‫ة‬ ‫قنواتنا‬ ‫على‬ ‫وادعمنا‬ ‫اإلنترنت‬ ‫على‬
‫على‬ ‫ملفي‬ ‫في‬ ‫هنا‬ ‫أرفعها‬ ‫التي‬ ‫شرائحي‬ ‫ومذاكرة‬ ‫بالتعلم‬ ‫وأسعد‬ ‫وأرحب‬ ‫أسمح‬ Slide share website ‫وال‬ ‫مانع‬ ‫أيضا‬ ‫من‬ ‫نسخ‬ ‫شريحة‬ ‫او‬ ‫اثنين‬ ‫عند‬ ‫الضرورة‬ ، ‫وال‬ ‫مانع‬ ‫من‬ ‫شرح‬ ‫البورب‬ ‫وينت‬ ‫الخاصة‬ ‫بي‬ ‫للغير‬ ‫بشرط‬ ‫عدم‬ ‫إزالة‬ ‫اسمي‬ ‫من‬ ‫البوربوينت‬ ، ‫فال‬ ‫أسمح‬ ‫أبدا‬ ‫بإزال‬ ‫ة‬ ‫اسمي‬ ‫من‬ ‫على‬ ‫الباوربوينت‬ ‫ووضع‬ ‫اسمك‬ ‫بدال‬ ‫منه‬ ‫لتصبح‬ ‫وكأنك‬ ‫من‬ ‫صممتها‬ ‫فهذه‬ ‫سرقة‬ ‫ال‬ ‫أسمح‬ ‫بها‬ ‫وتضييع‬ ‫لحق‬ ‫من‬ ‫تعب‬ ‫في‬ ‫عملها‬ . ‫وفقكم‬ ‫هللا‬ ‫وإياي‬ ‫للتعلم‬ ‫ونفع‬ ‫اآلخرين‬ I allow, welcome, and be happy to learn and study my slides that I upload here in my profile on Slide share website There is also no objection to copying one or two slides when necessary, and there is no objection to explaining my PowerPoint to others on the condition that my name is not removed from the PowerPoint. I never allow my name to be removed from PowerPoint and to replace it with yours, Make it look like you designed it. This is theft that I do not allow and a waste of the right of those who are tired in this work. May God bless you and me for learning and benefiting others 5
1- Cell membrane and nuclear membrane lysing. 2- Protein removal. 3- RNA degradation. 4- DNA precipitation. 5-Concentration and purity measuring. 4
DNA extraction ‫بسم‬ ‫هللا‬ ‫الرحمن‬ ‫الرحيم‬ ‫هتكلم‬ ‫اليوم‬ ‫إن‬ ‫شاء‬ ‫هللا‬ ‫عن‬ DNA extraction ‫بنجيب‬ ‫عينة‬ ‫الدم‬ 200 micron of whole blood ‫ونحط‬ ‫عليه‬ 20 micron proteinase K ‫ده‬ ‫هيكسر‬ ‫كل‬ ‫بروتينات‬ ‫الخاليا‬ 7
DNA extraction ‫نعمل‬ mix by vortex ‫ثم‬ ‫نضع‬ ‫عليه‬ 400 micron cell lysis solution ‫هيكسر‬ ‫ال‬ cell membrane ‫وغيره‬ ‫نعمل‬ mix by vortex ‫نترك‬ ‫هذا‬ ‫الخليط‬ 10 ‫دقائق‬ ‫في‬ ‫درجة‬ ‫حرارة‬ 56 ‫يبقي‬ ‫كده‬ ‫كسرت‬ ‫البروتينات‬ ‫الموجودة‬ ‫داخل‬ ‫الخلية‬ ‫وك‬ ‫سرت‬ ‫جدار‬ ‫الخاليا‬ ‫بعد‬ ‫ذلك‬ ‫نضع‬ 200 micron Ethanol to precipitate DNA DNA extraction 8
‫بعد‬ ‫ذلك‬ ‫ننقل‬ ‫هذا‬ ‫الخليط‬ ‫في‬ spin column ‫ونعمل‬ centrifugation ‫بمعدل‬ 7000 rpm ‫لمدة‬ ‫دقيقة‬ ‫كل‬ ‫البروتينات‬ ‫والدهون‬ ‫وأي‬ ‫شيء‬ ‫غير‬ ‫ال‬ DNA ‫هيتصفي‬ ‫في‬ ‫األنبوبة‬ ‫أسفل‬ ‫ال‬ spin column ‫وال‬ DNA ‫هيمسك‬ ‫في‬ silica ‫اللي‬ ‫داخل‬ ‫ال‬ spin column ‫يبقي‬ ‫كده‬ ‫تخلصنا‬ ‫من‬ ‫كل‬ ‫شيء‬ ‫باستثناء‬ ‫ال‬ DNA ‫واشياء‬ ‫صغيرة‬ ‫عالقة‬ ‫به‬ ‫ونرمي‬ ‫ما‬ ‫جمع‬ ‫في‬ tube ‫التي‬ ‫باألسفل‬ DNA extraction 9
DNA extraction ‫نضع‬ ‫ذلك‬ ‫بعد‬ • 500 micron of wash I ‫بال‬ ‫عالق‬ ‫أي‬ ‫نشيل‬ ‫علشان‬ DNA ‫ونعمل‬ centrifugation ‫بمعدل‬ 8000 rpm ‫في‬ ‫جمع‬ ‫ما‬ ‫ونرمي‬ ‫دقيقة‬ ‫لمدة‬ tube ‫نضع‬ ‫ذلك‬ ‫بعد‬ • 500 micron of wash II ‫بال‬ ‫عالق‬ ‫أي‬ ‫نشيل‬ ‫علشان‬ DNA ‫ونعمل‬ centrifugation ‫بمعدل‬ 20000 rpm ‫لمدة‬ 3 ‫في‬ ‫جمع‬ ‫ما‬ ‫ونرمي‬ ‫دقائق‬ tube ‫ال‬ ‫أن‬ ‫من‬ ‫للتأكد‬ ‫دي‬ ‫الخطوة‬ ‫نكرر‬ ‫ممكن‬ silica ‫شيء‬ ‫أي‬ ‫تحمل‬ ‫تعد‬ ‫لم‬ ‫ال‬ ‫سوي‬ DNA 10
DNA extraction ‫بعد‬ ‫ذلك‬ ‫نضع‬ 200 micron of ellution ‫فى‬ ‫منتصف‬ ‫األنبوبة‬ ‫لكي‬ ‫يمتز‬ ‫ا‬ ‫ل‬ DNA ‫اللي‬ ‫موجود‬ ‫علي‬ silica ‫وينزل‬ ‫في‬ ‫ال‬ tube ‫التي‬ ‫باألسفل‬ ‫ونعمل‬ centrifugation ‫بمعدل‬ 10000 rpm ‫لمدة‬ ‫دقيقتين‬ ‫هينزل‬ ‫في‬ ‫أسفل‬ ‫األنبوبة‬ ‫ال‬ DNA ‫أحتفظ‬ ‫بيه‬ ‫عند‬ ‫درجة‬ - 20 ‫لمدة‬ ‫شهور‬ ‫ودرجة‬ - 80 ‫لمدة‬ ‫سنين‬ ‫لحين‬ ‫استخدامه‬ . 11
‫بعد‬ ‫كده‬ ‫علشان‬ ‫نتأكد‬ ‫من‬ ‫إن‬ Purity of DNA ‫بنقيس‬ ‫ال‬ absorption in wl 260 and 280 by nanodrop ‫ونقسمهم‬ ‫علي‬ ‫بعض‬ ‫لو‬ ‫طلع‬ ‫بين‬ 1.8 - 2.1 ‫يبقي‬ ‫كده‬ ‫احنا‬ ‫ماشيين‬ ‫صح‬ . ‫وممكن‬ ‫نشوف‬ ‫احنا‬ ‫ماشيين‬ ‫صح‬ ‫وال‬ ‫غلط‬ ‫عن‬ ‫طريق‬ ‫ال‬ electrophoresis ‫أالقي‬ sharp band ‫في‬ ‫مكان‬ ‫انا‬ ‫متوقعه‬ ‫ومفيش‬ ‫أي‬ bands ‫أخري‬ nonspecific ‫كده‬ ‫أعرف‬ ‫اني‬ ‫ماشي‬ ‫صح‬ . ‫وممكن‬ ‫نفيس‬ ‫ال‬ concentration of DNA 12
13
DNA analysis by agarose gel electrophoresis. 14
RNA extraction 1 _ ‫ال‬ ‫خطوتي‬ ‫لعمل‬ ‫داعي‬ ‫فال‬ ‫السيروم‬ ‫في‬ ‫فيروس‬ ‫بفصل‬ ‫أنا‬ ‫لو‬ ‫طب‬ Proteinase K and cell lysis . ٢ _ ‫أفصل‬ ‫عاوز‬ ‫لو‬ ‫طبعا‬ RNA ‫أحط‬ ‫ما‬ ‫بدل‬ ‫بس‬ ‫الكالم‬ ‫نفس‬ ‫هو‬ RNAs ‫ال‬ ‫يكسر‬ RNA ‫أحط‬ DNAs ‫ال‬ ‫يكسر‬ DNA ‫وال‬ RNA ‫هو‬ ‫كما‬ ‫يظل‬ . ٣ _ ‫بستخلص‬ ‫أنا‬ ‫لو‬ RNA ‫منه‬ ‫أجيب‬ ‫هضطر‬ cDNA ‫ال‬ ‫جهاز‬ ‫ألن‬ PCR ‫ال‬ ‫علي‬ ‫إال‬ ‫يعمل‬ ‫ال‬ DNA ‫يسمي‬ ‫مشهور‬ ‫بإنزيم‬ ‫وذلك‬ Reverse Transcriptase If RNA is extracted, a reverse transcription step is required to form cDNA before amplification of DNA by PCR is possible. 15
RNA extraction ‫ال‬ ‫بستخلص‬ ‫وليه‬ ‫امتي‬ ‫طب‬ ‫المهم‬ ‫السؤال‬ RNA ‫؟‬ ‫حالتين‬ ‫في‬ : 1- detection of RNA viruses as HCV,HIV 2- detect gene expression ‫ل‬ ‫اترجم‬ ‫الجين‬ ‫اشوف‬ ‫عاوز‬ ‫يعني‬ mRNA ‫بروتين‬ ‫وهيطلع‬ ‫شغال‬ ‫ومش‬ ‫موجود‬ ‫هوه‬ ‫وال‬ ،‫ويشتغل‬ Not expressed. 16
DNA,RNA extraction • 20 µL proteinase K to 200 µl whole blood, Mix by vortex • Add 400 µL of lysis solution , mix by vortex to obtain uniform suspension incubate at 56 for 10 min • Add 200µl of ethanol (96-100%) and mix. • Transfere the prepared mixture to spin column . Centrifuge for 1 min at 8000 rpm. • Discard the collection tube containing the flow throw solution • Place column into a new 2 ml collection tube • Add 500 µl of wash buffer I and centrifuge for 1 min at 8000 rpm Discard the collection tube containing the flow throw solution place the column back into the collection tube • Add 500 µl of wash buffer II and centrifuge for 3 min at 14 000 rpm Discard the collection tube containing the flow throw solution place the column back into the collection tube. 17
DNA,RNA extraction • Empty the collection tube . Place the purification column back into the tube and re-spin the column for one min at maximum speed <14.000 rpm Discard the collection tube containing the flow throw solution • Place column into a new 1.5 ml collection tube • Add 200 µl of elution buffer to center of the column membrane to elute genomic DNA . Incubate 2 min at room temperature and centrifuge for I min at 10.000 rpm • Discard the prification column to be used immediately or stored at -20. 18
19
REFERENCES : • _ https://labtestsonline.org • _ https://www.medscape.com • _ https://www.wikipedia.org • _ https ps://www.labcorp.com • _ https://www.uptodate.com • _ https://www.ncbi.nlm.nih.gov Home - PubMed – NCBI • _TIETZ TEXTBOOK OF CLINICAL CHEMISTRY AND MOLECULAR DIAGNOSTICS, SIXTH EDITION 2018. • _Essential of clinical pathology book; 1st edition; Shirish M Kawthalkar; 2010. • _Essential of biochemistry book ;1st edition; 2012. • _ ABC of clinical hematology ; 4th edition ; Drew Provan; 2018. • _ Harper's illustrated biochemistry 30th edition 2015. • _ Lippincott's illustrated review of biochemistry sixth edition 2014. • _ Lecturea Notes Clinical Biochemistry, 9th Edition Walker, Simon, 2103. • _Some audios and videos from Well-known, trusted professors who study from accredited books. 20
‫اليوتيوب‬ ‫علي‬ ‫البحث‬ ‫أو‬ ،‫اليوتيوب‬ ‫علي‬ ‫قناتي‬ ‫دي‬ ‫د‬ ‫باسم‬ . ‫سفيان‬ ‫بهجت‬ ‫محمد‬ ، ‫وتشتركو‬ ‫تتابعوني‬ ‫ياريت‬ ‫حتى‬ ‫بالقناة‬ ‫ا‬ ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ https://www.youtube.com/channel/UCaYs1d8s0ntZvteHS3mMmGA ‫ب‬ ‫الفيس‬ ‫في‬ ‫البحث‬ ‫أو‬ ،‫انضمامكم‬ ‫يشرفنا‬ ‫الفيسبوك‬ ‫علي‬ ‫بتاعتي‬ ‫الصفحة‬ ‫دي‬ DMBMS2018 ، ‫حتى‬ ‫تتابعوني‬ ‫ياريت‬ ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ https://www.facebook.com/DMBMS2018/ ‫باسم‬ ‫التلجرام‬ ‫علي‬ ‫قناتي‬ ‫ودي‬ ِ ‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ 📚 ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ ‫حتى‬ ‫تتابعوني‬ ‫ِريت‬‫ا‬‫ي‬ https://t.me/DMBMS2020 ‫على‬ ‫بروفايلي‬ ‫لينكيدإن‬ ‫وتعملولي‬ ‫تتابعوني‬ ‫ياريت‬ endorsement ‫مهاراتي‬ ‫على‬ https://www.linkedin.com/in/mohammed-bahgat-sofyan-8ba012142/ ‫موقع‬ ‫على‬ ‫بروفايلي‬ ‫وده‬ SlideShare ‫تتابعونا‬ ‫ياريت‬ https://www.slideshare.net/MohammedBahgatMohamm1 ‫وده‬ ‫بروفايلي‬ ‫الشخصي‬ ‫الفيسبوك‬ ‫علي‬ https://www.facebook.com/mohammed.bahgat.165 ‫الفيسبوك‬ ‫على‬ ‫بنا‬ ‫الخاص‬ ‫اللوجوا‬ ‫وده‬ # ‫دمحمد‬ _ ‫بهجت‬ _ ‫سفيان‬ _ medical_biochimestry 21
OUR PAGES, CHANNELS AND GROUPS ON THE INTERNET ‫ونستفي‬ ‫عليهم‬ ‫البوستات‬ ‫كل‬ ‫نضع‬ ‫هللا‬ ‫شاء‬ ‫ان‬ ‫جميعا‬ ‫د‬ ‫انضمامكم‬ ‫يشرفنا‬ ‫وطبعا‬ • ‫إن‬ ‫شاء‬ ‫هللا‬ ‫يادكاترة‬ ‫الصفحة‬ ‫دي‬ ‫والقناة‬ ‫دي‬ ‫هتفيدكم‬ ،‫جدا‬ ‫مهتمية‬ ‫بالتحاليل‬ ‫الطبية‬ ‫هتف‬ ‫يد‬ ‫كل‬ ‫التخصصات‬ ‫الطبية‬ ‫ان‬ ‫شاء‬ ‫هللا‬ ‫وعليها‬ ‫برده‬ ‫بوستات‬ ‫دينية‬ ‫وأدبية‬ ‫يارب‬ ‫ينفع‬ ‫بيها‬ ‫ي‬ ‫ارب‬ ‫ويجعلها‬ ‫خالصة‬ ‫لوجهه‬ . https://www.youtube.com/channel/UCaYs1d8s0ntZvteHS3mMmGA • ‫د‬ ‫باسم‬ ‫اليوتيوب‬ ‫علي‬ ‫البحث‬ ‫أو‬ ،‫اليوتيوب‬ ‫علي‬ ‫قناتي‬ . ‫سفيان‬ ‫بهجت‬ ‫محمد‬ . • https://www.facebook.com/DMBMS2018/ • ‫ب‬ ‫الفيس‬ ‫في‬ ‫البحث‬ ‫أو‬ ،‫انضمامكم‬ ‫يشرفنا‬ ‫الفيسبوك‬ ‫علي‬ ‫بتاعتي‬ ‫الصفحة‬ ‫دي‬ DMBMS2018 • https://t.me/DMBMS2020 • ‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ ‫باسم‬ ‫التلجرام‬ ‫علي‬ ‫قناتي‬ ‫ودي‬ 📚 • ‫الدخو‬ ‫يرجي‬ ‫جروبين‬ ،‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ ‫باسم‬ ‫الواتس‬ ‫علي‬ ‫بي‬ ‫الخاص‬ ‫الجروب‬ ‫وده‬ ‫علي‬ ‫ل‬ ‫واحد‬ ‫المحتوي‬ ‫ألن‬ ‫فقط‬ ‫واحد‬ • https://chat.whatsapp.com/9vsZpHhVScfAe43xm8vBzm • https://chat.whatsapp.com/DPtwIYJlBJo4cTZnZ0OPir • ‫الفيسبوك‬ ‫علي‬ ‫الشخصي‬ ‫بروفايلي‬ ‫وده‬ • https://www.facebook.com/mohammed.bahgat.165 . • ‫الفيسبوك‬ ‫على‬ ‫بنا‬ ‫الخاص‬ ‫اللوجوا‬ ‫وده‬ # ‫دمحمد‬ _ ‫بهجت‬ _ ‫سفيان‬ _ medical_biochimestry 22
T H A N K Y O U M E E T O N T H E B E S T L A T E R , G O D W I L L I N G 23

Recommended

DNA fingerprint
DNA fingerprintDNA fingerprint
DNA fingerprint
Mohammed Bahgat Mohammed Sofyan
 
Mutation
MutationMutation
Mutation
Mohammed Bahgat Mohammed Sofyan
 
Calculation
CalculationCalculation
Calculation
Mohammed Bahgat Mohammed Sofyan
 
Calculation
CalculationCalculation
Calculation
Mohammed Bahgat Mohammed Sofyan
 
Mutation
MutationMutation
Mutation
Mohammed Bahgat Sofyan
 
13. Genetic Fingerprint.ppsx
13. Genetic Fingerprint.ppsx13. Genetic Fingerprint.ppsx
13. Genetic Fingerprint.ppsx
SakhiratulLaila
 
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docxPost-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
ChantellPantoja184
 
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docxHBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
P.B.PRAVEEN KUMAR
 

Recommended

DNA fingerprint
DNA fingerprintDNA fingerprint
DNA fingerprint
Mohammed Bahgat Mohammed Sofyan
 
Mutation
MutationMutation
Mutation
Mohammed Bahgat Mohammed Sofyan
 
Calculation
CalculationCalculation
Calculation
Mohammed Bahgat Mohammed Sofyan
 
Calculation
CalculationCalculation
Calculation
Mohammed Bahgat Mohammed Sofyan
 
Mutation
MutationMutation
Mutation
Mohammed Bahgat Sofyan
 
13. Genetic Fingerprint.ppsx
13. Genetic Fingerprint.ppsx13. Genetic Fingerprint.ppsx
13. Genetic Fingerprint.ppsx
SakhiratulLaila
 
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docxPost-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
Post-lab 1- Myths in Science (10 pts)Read the remaining myths” .docx
ChantellPantoja184
 
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docxHBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
HBV DNA QUANTITATIVE PCR UG ELECTIVE POSTINGS.docx
P.B.PRAVEEN KUMAR
 
Himanshu chaudhry
Himanshu chaudhryHimanshu chaudhry
Himanshu chaudhry
Himanshu Chaudhry
 
DNA Extraction, DNA quantity-quality check & Amplicon quantity check
DNA Extraction, DNA quantity-quality check & Amplicon quantity checkDNA Extraction, DNA quantity-quality check & Amplicon quantity check
DNA Extraction, DNA quantity-quality check & Amplicon quantity check
Mohiuddin Masum
 
Acknowledgments
AcknowledgmentsAcknowledgments
Acknowledgments
Mohsin Shad
 
RNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptxRNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptx
ASJADRAZA10
 
Purification of nucleic acids
Purification of nucleic acidsPurification of nucleic acids
Purification of nucleic acids
Yashswee Ghorpade
 
Steroid radioimmunoassay
Steroid radioimmunoassaySteroid radioimmunoassay
Steroid radioimmunoassay
Teuku Salman
 
enzyme (2)
enzyme (2)enzyme (2)
enzyme (2)
Sapna Singh
 
Training report at Chandigarh University
Training report at Chandigarh University Training report at Chandigarh University
Training report at Chandigarh University
IshantGupta58
 
JBK Pipetting SOP and Procedure
JBK Pipetting SOP and ProcedureJBK Pipetting SOP and Procedure
JBK Pipetting SOP and Procedure
John Kelley
 
B2.4 extracting dna
B2.4 extracting dnaB2.4 extracting dna
B2.4 extracting dna
Miss Lavin
 
الإيمان بالله جل وعلا.pptx
الإيمان بالله جل وعلا.pptxالإيمان بالله جل وعلا.pptx
الإيمان بالله جل وعلا.pptx
Mohammed Bahgat Mohammed Sofyan
 
Tests for kidney functions in blood
Tests for kidney functions in bloodTests for kidney functions in blood
Tests for kidney functions in blood
Mohammed Bahgat Mohammed Sofyan
 
Cholesterol
CholesterolCholesterol
Cholesterol
Mohammed Bahgat Mohammed Sofyan
 
Conventional , qPCR & RTPCR
Conventional , qPCR & RTPCRConventional , qPCR & RTPCR
Conventional , qPCR & RTPCR
Mohammed Bahgat Mohammed Sofyan
 
Ketonuria
KetonuriaKetonuria
Ketonuria
Mohammed Bahgat Mohammed Sofyan
 
Interpretation of ketoacidosis (dka) part 2
Interpretation of ketoacidosis (dka) part 2Interpretation of ketoacidosis (dka) part 2
Interpretation of ketoacidosis (dka) part 2
Mohammed Bahgat Mohammed Sofyan
 
Protein Electrophoresis
Protein ElectrophoresisProtein Electrophoresis
Protein Electrophoresis
Mohammed Bahgat Mohammed Sofyan
 
Protein Electrophoresis 2 nd year student
Protein Electrophoresis 2 nd year student Protein Electrophoresis 2 nd year student
Protein Electrophoresis 2 nd year student
Mohammed Bahgat Mohammed Sofyan
 
Chromatography
ChromatographyChromatography
Chromatography
Mohammed Bahgat Mohammed Sofyan
 
Total protein , globulin and a g ratio
Total protein , globulin and a g ratioTotal protein , globulin and a g ratio
Total protein , globulin and a g ratio
Mohammed Bahgat Mohammed Sofyan
 
Jaundice
JaundiceJaundice
Jaundice
Mohammed Bahgat Mohammed Sofyan
 
Albumin
AlbuminAlbumin
Albumin
Mohammed Bahgat Mohammed Sofyan
 

More Related Content

Similar to DNA & RNA extraction

RNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptxRNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptx
ASJADRAZA10
 
Purification of nucleic acids
Purification of nucleic acidsPurification of nucleic acids
Purification of nucleic acids
Yashswee Ghorpade
 
JBK Pipetting SOP and Procedure
JBK Pipetting SOP and ProcedureJBK Pipetting SOP and Procedure
JBK Pipetting SOP and Procedure
John Kelley
 
B2.4 extracting dna
B2.4 extracting dnaB2.4 extracting dna
B2.4 extracting dna
Miss Lavin
 

Similar to DNA & RNA extraction (10)

Himanshu chaudhry
Himanshu chaudhryHimanshu chaudhry
Himanshu chaudhry
 
DNA Extraction, DNA quantity-quality check & Amplicon quantity check
DNA Extraction, DNA quantity-quality check & Amplicon quantity checkDNA Extraction, DNA quantity-quality check & Amplicon quantity check
DNA Extraction, DNA quantity-quality check & Amplicon quantity check
 
Acknowledgments
AcknowledgmentsAcknowledgments
Acknowledgments
 
RNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptxRNA, DNA Isolation and cDNA synthesis.pptx
RNA, DNA Isolation and cDNA synthesis.pptx
 
Purification of nucleic acids
Purification of nucleic acidsPurification of nucleic acids
Purification of nucleic acids
 
Steroid radioimmunoassay
Steroid radioimmunoassaySteroid radioimmunoassay
Steroid radioimmunoassay
 
enzyme (2)
enzyme (2)enzyme (2)
enzyme (2)
 
Training report at Chandigarh University
Training report at Chandigarh University Training report at Chandigarh University
Training report at Chandigarh University
 
JBK Pipetting SOP and Procedure
JBK Pipetting SOP and ProcedureJBK Pipetting SOP and Procedure
JBK Pipetting SOP and Procedure
 
B2.4 extracting dna
B2.4 extracting dnaB2.4 extracting dna
B2.4 extracting dna
 

More from Mohammed Bahgat Mohammed Sofyan

More from Mohammed Bahgat Mohammed Sofyan (15)

الإيمان بالله جل وعلا.pptx
الإيمان بالله جل وعلا.pptxالإيمان بالله جل وعلا.pptx
الإيمان بالله جل وعلا.pptx
 
Tests for kidney functions in blood
Tests for kidney functions in bloodTests for kidney functions in blood
Tests for kidney functions in blood
 
Cholesterol
CholesterolCholesterol
Cholesterol
 
Conventional , qPCR & RTPCR
Conventional , qPCR & RTPCRConventional , qPCR & RTPCR
Conventional , qPCR & RTPCR
 
Ketonuria
KetonuriaKetonuria
Ketonuria
 
Interpretation of ketoacidosis (dka) part 2
Interpretation of ketoacidosis (dka) part 2Interpretation of ketoacidosis (dka) part 2
Interpretation of ketoacidosis (dka) part 2
 
Protein Electrophoresis
Protein ElectrophoresisProtein Electrophoresis
Protein Electrophoresis
 
Protein Electrophoresis 2 nd year student
Protein Electrophoresis 2 nd year student Protein Electrophoresis 2 nd year student
Protein Electrophoresis 2 nd year student
 
Chromatography
ChromatographyChromatography
Chromatography
 
Total protein , globulin and a g ratio
Total protein , globulin and a g ratioTotal protein , globulin and a g ratio
Total protein , globulin and a g ratio
 
Jaundice
JaundiceJaundice
Jaundice
 
Albumin
AlbuminAlbumin
Albumin
 
Hb electrophoresis
Hb electrophoresisHb electrophoresis
Hb electrophoresis
 
Curriculum of Junior medical staff online teaching
Curriculum of Junior medical staff online teachingCurriculum of Junior medical staff online teaching
Curriculum of Junior medical staff online teaching
 
أركان الإيمان وعقيدة المسلم
أركان الإيمان وعقيدة المسلم أركان الإيمان وعقيدة المسلم
أركان الإيمان وعقيدة المسلم
 

Recently uploaded

Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
EADTU
 
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdf
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdfContoh Aksi Nyata Refleksi Diri ( NUR ).pdf
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdf
cupulin
 
Spellings Wk 4 and Wk 5 for Grade 4 at CAPS
Spellings Wk 4 and Wk 5 for Grade 4 at CAPSSpellings Wk 4 and Wk 5 for Grade 4 at CAPS
Spellings Wk 4 and Wk 5 for Grade 4 at CAPS
AnaAcapella
 
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
中 央社
 
SURVEY I created for uni project research
SURVEY I created for uni project researchSURVEY I created for uni project research
SURVEY I created for uni project research
CaitlinCummins3
 
Personalisation of Education by AI and Big Data - Lourdes Guàrdia
Personalisation of Education by AI and Big Data - Lourdes GuàrdiaPersonalisation of Education by AI and Big Data - Lourdes Guàrdia
Personalisation of Education by AI and Big Data - Lourdes Guàrdia
EADTU
 

Recently uploaded (20)

How To Create Editable Tree View in Odoo 17
How To Create Editable Tree View in Odoo 17How To Create Editable Tree View in Odoo 17
How To Create Editable Tree View in Odoo 17
 
Graduate Outcomes Presentation Slides - English (v3).pptx
Graduate Outcomes Presentation Slides - English (v3).pptxGraduate Outcomes Presentation Slides - English (v3).pptx
Graduate Outcomes Presentation Slides - English (v3).pptx
 
Trauma-Informed Leadership - Five Practical Principles
Trauma-Informed Leadership - Five Practical PrinciplesTrauma-Informed Leadership - Five Practical Principles
Trauma-Informed Leadership - Five Practical Principles
 
diagnosting testing bsc 2nd sem.pptx....
diagnosting testing bsc 2nd sem.pptx....diagnosting testing bsc 2nd sem.pptx....
diagnosting testing bsc 2nd sem.pptx....
 
Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
Transparency, Recognition and the role of eSealing - Ildiko Mazar and Koen No...
 
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
 
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdf
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdfContoh Aksi Nyata Refleksi Diri ( NUR ).pdf
Contoh Aksi Nyata Refleksi Diri ( NUR ).pdf
 
How to Manage Website in Odoo 17 Studio App.pptx
How to Manage Website in Odoo 17 Studio App.pptxHow to Manage Website in Odoo 17 Studio App.pptx
How to Manage Website in Odoo 17 Studio App.pptx
 
Spellings Wk 4 and Wk 5 for Grade 4 at CAPS
Spellings Wk 4 and Wk 5 for Grade 4 at CAPSSpellings Wk 4 and Wk 5 for Grade 4 at CAPS
Spellings Wk 4 and Wk 5 for Grade 4 at CAPS
 
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
TỔNG HỢP HƠN 100 ĐỀ THI THỬ TỐT NGHIỆP THPT TOÁN 2024 - TỪ CÁC TRƯỜNG, TRƯỜNG...
 
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文會考英文
 
Major project report on Tata Motors and its marketing strategies
Major project report on Tata Motors and its marketing strategiesMajor project report on Tata Motors and its marketing strategies
Major project report on Tata Motors and its marketing strategies
 
SURVEY I created for uni project research
SURVEY I created for uni project researchSURVEY I created for uni project research
SURVEY I created for uni project research
 
COMMUNICATING NEGATIVE NEWS - APPROACHES .pptx
COMMUNICATING NEGATIVE NEWS - APPROACHES .pptxCOMMUNICATING NEGATIVE NEWS - APPROACHES .pptx
COMMUNICATING NEGATIVE NEWS - APPROACHES .pptx
 
AIM of Education-Teachers Training-2024.ppt
AIM of Education-Teachers Training-2024.pptAIM of Education-Teachers Training-2024.ppt
AIM of Education-Teachers Training-2024.ppt
 
Personalisation of Education by AI and Big Data - Lourdes Guàrdia
Personalisation of Education by AI and Big Data - Lourdes GuàrdiaPersonalisation of Education by AI and Big Data - Lourdes Guàrdia
Personalisation of Education by AI and Big Data - Lourdes Guàrdia
 
OS-operating systems- ch05 (CPU Scheduling) ...
OS-operating systems- ch05 (CPU Scheduling) ...OS-operating systems- ch05 (CPU Scheduling) ...
OS-operating systems- ch05 (CPU Scheduling) ...
 
FICTIONAL SALESMAN/SALESMAN SNSW 2024.pdf
FICTIONAL SALESMAN/SALESMAN SNSW 2024.pdfFICTIONAL SALESMAN/SALESMAN SNSW 2024.pdf
FICTIONAL SALESMAN/SALESMAN SNSW 2024.pdf
 
Basic Civil Engineering notes on Transportation Engineering & Modes of Transport
Basic Civil Engineering notes on Transportation Engineering & Modes of TransportBasic Civil Engineering notes on Transportation Engineering & Modes of Transport
Basic Civil Engineering notes on Transportation Engineering & Modes of Transport
 
Stl Algorithms in C++ jjjjjjjjjjjjjjjjjj
Stl Algorithms in C++ jjjjjjjjjjjjjjjjjjStl Algorithms in C++ jjjjjjjjjjjjjjjjjj
Stl Algorithms in C++ jjjjjjjjjjjjjjjjjj
 

DNA & RNA extraction

  • 1. DNA,RNA extraction BY dr: Mohammed Bahgat Mohammed Sofyan Assistant lecturer of medical biochemistry, Faculty of medicine, Al-Azhar university (Assiut branch) 1
  • 2. 2
  • 4. ‫صورة‬ ‫كل‬ ‫على‬ ‫اضغط‬ ‫الشاش‬ ‫عرض‬ ‫وضع‬ ‫في‬ ‫ة‬ ‫قنواتنا‬ ‫على‬ ‫وادعمنا‬ ‫اإلنترنت‬ ‫على‬
  • 5. ‫على‬ ‫ملفي‬ ‫في‬ ‫هنا‬ ‫أرفعها‬ ‫التي‬ ‫شرائحي‬ ‫ومذاكرة‬ ‫بالتعلم‬ ‫وأسعد‬ ‫وأرحب‬ ‫أسمح‬ Slide share website ‫وال‬ ‫مانع‬ ‫أيضا‬ ‫من‬ ‫نسخ‬ ‫شريحة‬ ‫او‬ ‫اثنين‬ ‫عند‬ ‫الضرورة‬ ، ‫وال‬ ‫مانع‬ ‫من‬ ‫شرح‬ ‫البورب‬ ‫وينت‬ ‫الخاصة‬ ‫بي‬ ‫للغير‬ ‫بشرط‬ ‫عدم‬ ‫إزالة‬ ‫اسمي‬ ‫من‬ ‫البوربوينت‬ ، ‫فال‬ ‫أسمح‬ ‫أبدا‬ ‫بإزال‬ ‫ة‬ ‫اسمي‬ ‫من‬ ‫على‬ ‫الباوربوينت‬ ‫ووضع‬ ‫اسمك‬ ‫بدال‬ ‫منه‬ ‫لتصبح‬ ‫وكأنك‬ ‫من‬ ‫صممتها‬ ‫فهذه‬ ‫سرقة‬ ‫ال‬ ‫أسمح‬ ‫بها‬ ‫وتضييع‬ ‫لحق‬ ‫من‬ ‫تعب‬ ‫في‬ ‫عملها‬ . ‫وفقكم‬ ‫هللا‬ ‫وإياي‬ ‫للتعلم‬ ‫ونفع‬ ‫اآلخرين‬ I allow, welcome, and be happy to learn and study my slides that I upload here in my profile on Slide share website There is also no objection to copying one or two slides when necessary, and there is no objection to explaining my PowerPoint to others on the condition that my name is not removed from the PowerPoint. I never allow my name to be removed from PowerPoint and to replace it with yours, Make it look like you designed it. This is theft that I do not allow and a waste of the right of those who are tired in this work. May God bless you and me for learning and benefiting others 5
  • 6. 1- Cell membrane and nuclear membrane lysing. 2- Protein removal. 3- RNA degradation. 4- DNA precipitation. 5-Concentration and purity measuring. 4
  • 7. DNA extraction ‫بسم‬ ‫هللا‬ ‫الرحمن‬ ‫الرحيم‬ ‫هتكلم‬ ‫اليوم‬ ‫إن‬ ‫شاء‬ ‫هللا‬ ‫عن‬ DNA extraction ‫بنجيب‬ ‫عينة‬ ‫الدم‬ 200 micron of whole blood ‫ونحط‬ ‫عليه‬ 20 micron proteinase K ‫ده‬ ‫هيكسر‬ ‫كل‬ ‫بروتينات‬ ‫الخاليا‬ 7
  • 8. DNA extraction ‫نعمل‬ mix by vortex ‫ثم‬ ‫نضع‬ ‫عليه‬ 400 micron cell lysis solution ‫هيكسر‬ ‫ال‬ cell membrane ‫وغيره‬ ‫نعمل‬ mix by vortex ‫نترك‬ ‫هذا‬ ‫الخليط‬ 10 ‫دقائق‬ ‫في‬ ‫درجة‬ ‫حرارة‬ 56 ‫يبقي‬ ‫كده‬ ‫كسرت‬ ‫البروتينات‬ ‫الموجودة‬ ‫داخل‬ ‫الخلية‬ ‫وك‬ ‫سرت‬ ‫جدار‬ ‫الخاليا‬ ‫بعد‬ ‫ذلك‬ ‫نضع‬ 200 micron Ethanol to precipitate DNA DNA extraction 8
  • 9. ‫بعد‬ ‫ذلك‬ ‫ننقل‬ ‫هذا‬ ‫الخليط‬ ‫في‬ spin column ‫ونعمل‬ centrifugation ‫بمعدل‬ 7000 rpm ‫لمدة‬ ‫دقيقة‬ ‫كل‬ ‫البروتينات‬ ‫والدهون‬ ‫وأي‬ ‫شيء‬ ‫غير‬ ‫ال‬ DNA ‫هيتصفي‬ ‫في‬ ‫األنبوبة‬ ‫أسفل‬ ‫ال‬ spin column ‫وال‬ DNA ‫هيمسك‬ ‫في‬ silica ‫اللي‬ ‫داخل‬ ‫ال‬ spin column ‫يبقي‬ ‫كده‬ ‫تخلصنا‬ ‫من‬ ‫كل‬ ‫شيء‬ ‫باستثناء‬ ‫ال‬ DNA ‫واشياء‬ ‫صغيرة‬ ‫عالقة‬ ‫به‬ ‫ونرمي‬ ‫ما‬ ‫جمع‬ ‫في‬ tube ‫التي‬ ‫باألسفل‬ DNA extraction 9
  • 10. DNA extraction ‫نضع‬ ‫ذلك‬ ‫بعد‬ • 500 micron of wash I ‫بال‬ ‫عالق‬ ‫أي‬ ‫نشيل‬ ‫علشان‬ DNA ‫ونعمل‬ centrifugation ‫بمعدل‬ 8000 rpm ‫في‬ ‫جمع‬ ‫ما‬ ‫ونرمي‬ ‫دقيقة‬ ‫لمدة‬ tube ‫نضع‬ ‫ذلك‬ ‫بعد‬ • 500 micron of wash II ‫بال‬ ‫عالق‬ ‫أي‬ ‫نشيل‬ ‫علشان‬ DNA ‫ونعمل‬ centrifugation ‫بمعدل‬ 20000 rpm ‫لمدة‬ 3 ‫في‬ ‫جمع‬ ‫ما‬ ‫ونرمي‬ ‫دقائق‬ tube ‫ال‬ ‫أن‬ ‫من‬ ‫للتأكد‬ ‫دي‬ ‫الخطوة‬ ‫نكرر‬ ‫ممكن‬ silica ‫شيء‬ ‫أي‬ ‫تحمل‬ ‫تعد‬ ‫لم‬ ‫ال‬ ‫سوي‬ DNA 10
  • 11. DNA extraction ‫بعد‬ ‫ذلك‬ ‫نضع‬ 200 micron of ellution ‫فى‬ ‫منتصف‬ ‫األنبوبة‬ ‫لكي‬ ‫يمتز‬ ‫ا‬ ‫ل‬ DNA ‫اللي‬ ‫موجود‬ ‫علي‬ silica ‫وينزل‬ ‫في‬ ‫ال‬ tube ‫التي‬ ‫باألسفل‬ ‫ونعمل‬ centrifugation ‫بمعدل‬ 10000 rpm ‫لمدة‬ ‫دقيقتين‬ ‫هينزل‬ ‫في‬ ‫أسفل‬ ‫األنبوبة‬ ‫ال‬ DNA ‫أحتفظ‬ ‫بيه‬ ‫عند‬ ‫درجة‬ - 20 ‫لمدة‬ ‫شهور‬ ‫ودرجة‬ - 80 ‫لمدة‬ ‫سنين‬ ‫لحين‬ ‫استخدامه‬ . 11
  • 12. ‫بعد‬ ‫كده‬ ‫علشان‬ ‫نتأكد‬ ‫من‬ ‫إن‬ Purity of DNA ‫بنقيس‬ ‫ال‬ absorption in wl 260 and 280 by nanodrop ‫ونقسمهم‬ ‫علي‬ ‫بعض‬ ‫لو‬ ‫طلع‬ ‫بين‬ 1.8 - 2.1 ‫يبقي‬ ‫كده‬ ‫احنا‬ ‫ماشيين‬ ‫صح‬ . ‫وممكن‬ ‫نشوف‬ ‫احنا‬ ‫ماشيين‬ ‫صح‬ ‫وال‬ ‫غلط‬ ‫عن‬ ‫طريق‬ ‫ال‬ electrophoresis ‫أالقي‬ sharp band ‫في‬ ‫مكان‬ ‫انا‬ ‫متوقعه‬ ‫ومفيش‬ ‫أي‬ bands ‫أخري‬ nonspecific ‫كده‬ ‫أعرف‬ ‫اني‬ ‫ماشي‬ ‫صح‬ . ‫وممكن‬ ‫نفيس‬ ‫ال‬ concentration of DNA 12
  • 13. 13
  • 14. DNA analysis by agarose gel electrophoresis. 14
  • 15. RNA extraction 1 _ ‫ال‬ ‫خطوتي‬ ‫لعمل‬ ‫داعي‬ ‫فال‬ ‫السيروم‬ ‫في‬ ‫فيروس‬ ‫بفصل‬ ‫أنا‬ ‫لو‬ ‫طب‬ Proteinase K and cell lysis . ٢ _ ‫أفصل‬ ‫عاوز‬ ‫لو‬ ‫طبعا‬ RNA ‫أحط‬ ‫ما‬ ‫بدل‬ ‫بس‬ ‫الكالم‬ ‫نفس‬ ‫هو‬ RNAs ‫ال‬ ‫يكسر‬ RNA ‫أحط‬ DNAs ‫ال‬ ‫يكسر‬ DNA ‫وال‬ RNA ‫هو‬ ‫كما‬ ‫يظل‬ . ٣ _ ‫بستخلص‬ ‫أنا‬ ‫لو‬ RNA ‫منه‬ ‫أجيب‬ ‫هضطر‬ cDNA ‫ال‬ ‫جهاز‬ ‫ألن‬ PCR ‫ال‬ ‫علي‬ ‫إال‬ ‫يعمل‬ ‫ال‬ DNA ‫يسمي‬ ‫مشهور‬ ‫بإنزيم‬ ‫وذلك‬ Reverse Transcriptase If RNA is extracted, a reverse transcription step is required to form cDNA before amplification of DNA by PCR is possible. 15
  • 16. RNA extraction ‫ال‬ ‫بستخلص‬ ‫وليه‬ ‫امتي‬ ‫طب‬ ‫المهم‬ ‫السؤال‬ RNA ‫؟‬ ‫حالتين‬ ‫في‬ : 1- detection of RNA viruses as HCV,HIV 2- detect gene expression ‫ل‬ ‫اترجم‬ ‫الجين‬ ‫اشوف‬ ‫عاوز‬ ‫يعني‬ mRNA ‫بروتين‬ ‫وهيطلع‬ ‫شغال‬ ‫ومش‬ ‫موجود‬ ‫هوه‬ ‫وال‬ ،‫ويشتغل‬ Not expressed. 16
  • 17. DNA,RNA extraction • 20 µL proteinase K to 200 µl whole blood, Mix by vortex • Add 400 µL of lysis solution , mix by vortex to obtain uniform suspension incubate at 56 for 10 min • Add 200µl of ethanol (96-100%) and mix. • Transfere the prepared mixture to spin column . Centrifuge for 1 min at 8000 rpm. • Discard the collection tube containing the flow throw solution • Place column into a new 2 ml collection tube • Add 500 µl of wash buffer I and centrifuge for 1 min at 8000 rpm Discard the collection tube containing the flow throw solution place the column back into the collection tube • Add 500 µl of wash buffer II and centrifuge for 3 min at 14 000 rpm Discard the collection tube containing the flow throw solution place the column back into the collection tube. 17
  • 18. DNA,RNA extraction • Empty the collection tube . Place the purification column back into the tube and re-spin the column for one min at maximum speed <14.000 rpm Discard the collection tube containing the flow throw solution • Place column into a new 1.5 ml collection tube • Add 200 µl of elution buffer to center of the column membrane to elute genomic DNA . Incubate 2 min at room temperature and centrifuge for I min at 10.000 rpm • Discard the prification column to be used immediately or stored at -20. 18
  • 19. 19
  • 20. REFERENCES : • _ https://labtestsonline.org • _ https://www.medscape.com • _ https://www.wikipedia.org • _ https ps://www.labcorp.com • _ https://www.uptodate.com • _ https://www.ncbi.nlm.nih.gov Home - PubMed – NCBI • _TIETZ TEXTBOOK OF CLINICAL CHEMISTRY AND MOLECULAR DIAGNOSTICS, SIXTH EDITION 2018. • _Essential of clinical pathology book; 1st edition; Shirish M Kawthalkar; 2010. • _Essential of biochemistry book ;1st edition; 2012. • _ ABC of clinical hematology ; 4th edition ; Drew Provan; 2018. • _ Harper's illustrated biochemistry 30th edition 2015. • _ Lippincott's illustrated review of biochemistry sixth edition 2014. • _ Lecturea Notes Clinical Biochemistry, 9th Edition Walker, Simon, 2103. • _Some audios and videos from Well-known, trusted professors who study from accredited books. 20
  • 21. ‫اليوتيوب‬ ‫علي‬ ‫البحث‬ ‫أو‬ ،‫اليوتيوب‬ ‫علي‬ ‫قناتي‬ ‫دي‬ ‫د‬ ‫باسم‬ . ‫سفيان‬ ‫بهجت‬ ‫محمد‬ ، ‫وتشتركو‬ ‫تتابعوني‬ ‫ياريت‬ ‫حتى‬ ‫بالقناة‬ ‫ا‬ ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ https://www.youtube.com/channel/UCaYs1d8s0ntZvteHS3mMmGA ‫ب‬ ‫الفيس‬ ‫في‬ ‫البحث‬ ‫أو‬ ،‫انضمامكم‬ ‫يشرفنا‬ ‫الفيسبوك‬ ‫علي‬ ‫بتاعتي‬ ‫الصفحة‬ ‫دي‬ DMBMS2018 ، ‫حتى‬ ‫تتابعوني‬ ‫ياريت‬ ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ https://www.facebook.com/DMBMS2018/ ‫باسم‬ ‫التلجرام‬ ‫علي‬ ‫قناتي‬ ‫ودي‬ ِ ‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ 📚 ‫هللا‬ ‫شاء‬ ‫إن‬ ‫نستمر‬ ‫حتى‬ ‫تتابعوني‬ ‫ِريت‬‫ا‬‫ي‬ https://t.me/DMBMS2020 ‫على‬ ‫بروفايلي‬ ‫لينكيدإن‬ ‫وتعملولي‬ ‫تتابعوني‬ ‫ياريت‬ endorsement ‫مهاراتي‬ ‫على‬ https://www.linkedin.com/in/mohammed-bahgat-sofyan-8ba012142/ ‫موقع‬ ‫على‬ ‫بروفايلي‬ ‫وده‬ SlideShare ‫تتابعونا‬ ‫ياريت‬ https://www.slideshare.net/MohammedBahgatMohamm1 ‫وده‬ ‫بروفايلي‬ ‫الشخصي‬ ‫الفيسبوك‬ ‫علي‬ https://www.facebook.com/mohammed.bahgat.165 ‫الفيسبوك‬ ‫على‬ ‫بنا‬ ‫الخاص‬ ‫اللوجوا‬ ‫وده‬ # ‫دمحمد‬ _ ‫بهجت‬ _ ‫سفيان‬ _ medical_biochimestry 21
  • 22. OUR PAGES, CHANNELS AND GROUPS ON THE INTERNET ‫ونستفي‬ ‫عليهم‬ ‫البوستات‬ ‫كل‬ ‫نضع‬ ‫هللا‬ ‫شاء‬ ‫ان‬ ‫جميعا‬ ‫د‬ ‫انضمامكم‬ ‫يشرفنا‬ ‫وطبعا‬ • ‫إن‬ ‫شاء‬ ‫هللا‬ ‫يادكاترة‬ ‫الصفحة‬ ‫دي‬ ‫والقناة‬ ‫دي‬ ‫هتفيدكم‬ ،‫جدا‬ ‫مهتمية‬ ‫بالتحاليل‬ ‫الطبية‬ ‫هتف‬ ‫يد‬ ‫كل‬ ‫التخصصات‬ ‫الطبية‬ ‫ان‬ ‫شاء‬ ‫هللا‬ ‫وعليها‬ ‫برده‬ ‫بوستات‬ ‫دينية‬ ‫وأدبية‬ ‫يارب‬ ‫ينفع‬ ‫بيها‬ ‫ي‬ ‫ارب‬ ‫ويجعلها‬ ‫خالصة‬ ‫لوجهه‬ . https://www.youtube.com/channel/UCaYs1d8s0ntZvteHS3mMmGA • ‫د‬ ‫باسم‬ ‫اليوتيوب‬ ‫علي‬ ‫البحث‬ ‫أو‬ ،‫اليوتيوب‬ ‫علي‬ ‫قناتي‬ . ‫سفيان‬ ‫بهجت‬ ‫محمد‬ . • https://www.facebook.com/DMBMS2018/ • ‫ب‬ ‫الفيس‬ ‫في‬ ‫البحث‬ ‫أو‬ ،‫انضمامكم‬ ‫يشرفنا‬ ‫الفيسبوك‬ ‫علي‬ ‫بتاعتي‬ ‫الصفحة‬ ‫دي‬ DMBMS2018 • https://t.me/DMBMS2020 • ‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ ‫باسم‬ ‫التلجرام‬ ‫علي‬ ‫قناتي‬ ‫ودي‬ 📚 • ‫الدخو‬ ‫يرجي‬ ‫جروبين‬ ،‫للناس‬ ‫أنفعهم‬ ‫الناس‬ ‫خير‬ ‫باسم‬ ‫الواتس‬ ‫علي‬ ‫بي‬ ‫الخاص‬ ‫الجروب‬ ‫وده‬ ‫علي‬ ‫ل‬ ‫واحد‬ ‫المحتوي‬ ‫ألن‬ ‫فقط‬ ‫واحد‬ • https://chat.whatsapp.com/9vsZpHhVScfAe43xm8vBzm • https://chat.whatsapp.com/DPtwIYJlBJo4cTZnZ0OPir • ‫الفيسبوك‬ ‫علي‬ ‫الشخصي‬ ‫بروفايلي‬ ‫وده‬ • https://www.facebook.com/mohammed.bahgat.165 . • ‫الفيسبوك‬ ‫على‬ ‫بنا‬ ‫الخاص‬ ‫اللوجوا‬ ‫وده‬ # ‫دمحمد‬ _ ‫بهجت‬ _ ‫سفيان‬ _ medical_biochimestry 22
  • 23. T H A N K Y O U M E E T O N T H E B E S T L A T E R , G O D W I L L I N G 23