lab and listening Flashcards
Concisely describe how to safely dispose the agar plates
Place in the plastic bag and autoclave
Heat melts the plastic agar plate
can be discarded
How to place the agar plate for incubation
upside down so that condensation doesn’t form on the side with the medium
37 degrees C and 5% CO2 atmosphere
Agar plates
petri dishes with an agar medium on it that allows for the growth of microorganisms
can have LB or antibiotics on it as well
LB
a broth created to promote bacterial growth
Ampicillin
an antibiotic that can be on the plate to help with the creation of a specific bacterial colony
antibiotic resistance
Permissive plate
allows the growth of any bacterial colony
Restrictive plate
allows growth of specific colonies based on antibiotic resistance
Indicator plate
shows colonies based on color
List the major procedures to making agar plates
Add 500 mL of distilled water into a 1 L flask
Insert LB agar tablets
cover with aluminum foil
autoclave
50 mg Amp and dissolve in 1 mL distilled water
LB flask cools to 55 degree C, add Amp mixture
Pour the liquid onto each plate (don’t open the lids all of the way) - about 5 mm
Swirl the plates in a circular motion, let cool for 20 min, incubate upside down
We have 60 uL digested DNA solution and 4X blue dye DNA loading buffer. In order to run DNA gel, we need to add a certain volume of 4X blue dye DNA loading buffer into DNA solution to obtain a final mixed solution with IX blue dye. Please calculate how much volume of 4X blue dye DNA loading buffer which you need to take and add it into DNA solution
60 + x = 4X
60 = 3X
20 = X
ゲ—ム
game
アリバイト
part time job
バイト
part time job
かいもの
shopping
クラス
class
いぬ
dog
ねこ
cat
ひと
person
こども
child
あなた
you
いす
chair
つくえ
desk
しゃしん
picture
はな
flower
レポ—ト
term paper
ごはん
rice
パン
bread
おてら
temple
こうえん
park
ス—パ—
supermarket
バスてい
bus stop
びょういん
hospital
ホテル
hotel
ほにゃ
bookstore
まち
town, city
レストラン
restaurant
geemu
game
aribaito
part time job
baito
part time job
kaimono
shopping
kurasu
class
inu
dog
neko
cat
hito
person
kodomo
child
anata
you
isu
chair
tsukue
desk
shashin
picture
hana
flower
repooto
term paper
gohan
rice, meal
pan
bread
otera
temple
kouen
park
suupaa
supermarket
basutei
bus stop
byouin
hospital
hoteru
hotel
honnya
bookstore
machi
town, city
resutoran
restaurant
We have 50 uL digested DNA solution and 5X blue dye DNA loading buffer. In order to run DNA gel, we need to add a certain volume of 5X blue dye DNA loading buffer into DNA solution to obtain a final mixed solution with 1X blue dye. Please calculate how much volume of 5X blue dye DNA loading buffer which you need to take and add it into DNA solution
50 + X = 5X
50 = 4x
12.5 = x
What do plasmids mean
Double-stranded
circular DNA
in bacteria
used for transformations of foreign DNA into bacterial cells
covalently closed
Are plasmids chromosomal molecules
No
extrachromosomal DNA that’s self-replicating
Vectors
used to carry exogenous DNA from somewhere else and insert it into a host cell
Plasmid vector
vectors where a plasmid is used to carry the exogenous DNA, so it’s transformed into recombinant DNA and introduced back into the cell
Competent cells
capable of taking exogenous DNA and have it inserted into their cells via transformation
What is the purpose to use competent cells
capable of taking exogenous DNA and have it inserted into their cells through a transformation
higher efficiency
used to propagate and maintain cloned DNA in plasmids
Transformation
exogenous DNA is taken into the new bacteria
replaces a section on the plasmid (Lac Z)
Major steps of a transformation
thaw competent cells
mix 2 uL sample and 50 uL competent cells
keep on ice for 30 min
heat shock 35 sec
ice 2 min
add LB
incubate on shaker at 37 degree C for 1 hour
plate on agar plate using sterile beads
incubate 37 degree C overnight
During transformation, which heat shock temperature is used
42 degree C
Why are heat shock time and temperature crucial
you don’t want to destroy your DNA or your cell
you want to heat it enough so that the DNA can enter the cell, but not too long that your bacteria cells begin to die
heat shock allows the DNA to enter through the membrane
What is the purpose to use the appropriate antibiotic during the transformation procedure
antibiotic resistance
transformed bacteria will be given an antibiotic resistance gene, so all non-transformed bacteria will die on the agar plate and the transformed bacteria will grow
Amp plate for Amp resistance
きのう
yesterday
…じかん
hour counter (duration)
いちじかん
for one hour
せんしゅう
last week
とき (の)
when…, at the time of…
げつようび
Monday
かようび
Tuesday
すいようび
Wednesday
もくようび
Thursday
きにょうい
Friday
あう (に)
to meet, to see (a person)
ある (に, が)
there is…
かう
to buy
かく
to write
とる
to take (a picture)
まつ
to wait
わかる (が)
to understand
いる (に, が)
there is… (living thing)
ぐらい
about (approximate measurement)
ごめんなさい
I’m sorry
それから
and then
だから
so, therefore
たくさん
many, a lot
と
together with (a person), and
どうして
why
ひとりで
alone
もし もし
hello? (on the phone)
kinou
yesterday
jikan
hour counter (duration)
ichijikan
for one hour
senshuu
last week
toki (no)
when…, at the time of…
getsuyoubi
monday
kayoubi
tuesday
suiyoubi
wednesday
mokuyoubi
thursday
kinyoubi
friday
au (ni)
to meet, to see (a person)
aru (ni, ga)
there is…
kau
to buy
kaku
to write
toru
to take (a picture)
matsu
to wait
wakaru (ga)
to understand
iru (ni, ga)
there is… (living thing)
gurai
about (approximate measurement)
gomennasai
i’m sorry
sorekara
and then
dakara
so, therefore
takusan
many, a lot
to
together with (a person), and
doushite
why
hitoride
alone
moshi moshi
hello? (on the phone)
What do restriction endonucleases or restriction enzymes mean
enzymes in bacteria created as a defense mechanism to digest foreign DNA
breaks DNA at a recognition sequence with 2 breaks (1 through backbone, 1 through double helix)
Recognition sequence
DNA-binding protein motif where restriction enzymes cut the DNA
blunt end
sticky end